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Credible and Facile Fluorometric Detection of Soybean Trypsin Inhibitor Activity with a Water-Soluble Poly(diphenylacetylene) Derivative
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作者 Jiao Tang Xiao Tang +7 位作者 Jiaxiang Tang Wei Qi Qianwei Pan Jinhong Zeng Housheng Xia Jianping Zhou Zhongyi Sheng Junfeng Niu 《Transactions of Tianjin University》 EI CAS 2024年第4期305-313,共9页
Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroani... Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroaniline,produced from the trypsin-catalyzed decomposition of N-benzoyl-DL-arginine-4-nitroanilide hydrochloride(L-BAPA),was well described using the Stern-Volmer equation.SBTI activity was quantitatively assessed based on changes in the fl uorescence intensity of the polymer.This strategy has several advantages,such as high sensitivity and ease of operation.Moreover,its applicability to other biochemical analyses is promising. 展开更多
关键词 Fluorometric detection Soybean trypsin inhibitor Inhibition activity Fluorescence quenching
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A Proteinase from Mung Bean Sprouts That Inactivaties Soybean Trypsin Inhibitor 被引量:2
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作者 陈中 杨晓泉 赵谋明 《Acta Botanica Sinica》 CSCD 2001年第11期1150-1153,共4页
By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE... By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h. 展开更多
关键词 PROTEINASE inactivate soybean trypsin inhibitor
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Optimization of Solid-State Fermentation with Lactobacillus brevis and Aspergillus oryzae for Trypsin Inhibitor Degradation in Soybean Meal 被引量:19
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作者 GAO You-ling WANG Cai-sheng +1 位作者 ZHU Qiu-hua QIAN Guo-ying 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第5期869-876,共8页
The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation (SSF) with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on ph... The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation (SSF) with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on phytic acid, crude protein, crude fat, and amino acid profile. Response surface methodology (RSM) with Box-Behnken design was used to optimize SSF. The optimal conditions derived from RSM for L. brevis fermentation were: pH=5. 1; inoculum size=10%; duration=72 h; substrate to water ratio=1.5. The minimum content of trypsin inhibitors was 6.4 mg g^-1 dry matter. The optimal conditions derived from RSM for A. oryzae fermentation were: substrate to water ratio= 0.8 1; inoculum size=4%; duration=120 h. The minimum content of trypsin inhibitors was 1.6 mg g^-1 dry matter. Both L. brevis and A. oryzae decreased trypsin inhibitors dramatically (57.1 and 89.2% respectively). L. brevis fermentation did not affect phytic acid (0.4%) and crude fat (5.2%) considerably, whereas A. oryzae fermentation degraded phytic acid (34.8%) and crude fat (22.0%) contents to a certain extent. Crude protein content was increased after both fermentation (6.4 and 12.9% for L. brevis and A. oryzae respectively). Urease activity was reduced greatly (83.3 and 58.3% for L. brevis and A. oryzae respectively). In conclusion, SSF with A. oryzae and L. brevis reduced trypsin inhibitor content and modified major macronutrients in soybean meal. 展开更多
关键词 Aspergillus oryzae Lactobacillus brevis response surface methodology solid-state fermentation soybean meal trypsin inhibitors
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Urinary trypsin inhibitor attenuates hepatic ischemia-reperfusion injury by reducing nuclear factor-kappa B activation 被引量:28
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作者 Wu, Yi-Jun Ling, Qi +4 位作者 Zhou, Xin-Hui Wang, Yan Xie, Hai-Yang Yu, Ji-Ren Zheng, Shu-Sen 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2009年第1期53-58,共6页
BACKGROUND: Urinary trypsin inhibitor (UTI) inhibits the inflammatory response and protects against ischemia-reperfusion (I/R) injury. The inflammatory response is mediated by nuclear factor-kappa B (NF-kappa B) and i... BACKGROUND: Urinary trypsin inhibitor (UTI) inhibits the inflammatory response and protects against ischemia-reperfusion (I/R) injury. The inflammatory response is mediated by nuclear factor-kappa B (NF-kappa B) and its related target genes and products such as vascular endothelial cell adhesion molecule and CXC chemokines. We aimed to assess the roles of those mediators in a UTI-treated mouse model of hepatic I/R injury. METHODS: Treatment group 1 (UTI given 5 minutes prior to liver ischemia), treatment group 2 (UTI given 5 minutes after the anhepatic phase) and a control group were investigated. Blood and liver samples were obtained and compared at 1, 3, 6 and 24 hours after reperfusion. RESULTS: Attenuation of pathological hepatocellular damage was greater in the treatment groups than in the control group (P < 0.05). Compared with the control group, the UTI treatment groups showed significantly lower serum alanine aminotransferase and aspartate aminotransferase levels, decreased myeloperoxidase activity, and reduced NF-kappa B activation. Also downregulated was the expression of tumor necrosis factor-alpha, cytokine-induced neutrophil chemoattractant, and macrophage inflammatory protein-2 at the mRNA level. P-selectin protein and intercellular adhesion molecule-1 protein expression were also downregulated. In addition, the treatment group I showed a better protective effect against I/R injury than the treatment group 2. CONCLUSIONS: UTI reduces NF-kappa B activation and downregulates the expression of its related mediators, followed by the inhibition of neutrophil aggregation and infiltration in hepatic I/R injury. The protective role of UTI is more effective in prevention than in treatment. 展开更多
关键词 ischemia-reperfusion injury nuclear factor-kappa B tumor necrosis factor-alpha urinary trypsin inhibitor
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Purification, characterization and evaluation of insecticidal activity of trypsin inhibitor from Albizia lebbeck seeds 被引量:1
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作者 Pratima Sharma Amarjit K Nath +1 位作者 Reena Kumari SV Bhardwaj 《Journal of Forestry Research》 CAS CSCD 2012年第1期131-137,共7页
A Bowman-Birk inhibitor with activity against gut proteases of Helicoverpa armigera was extracted in 0.I M sodium phosphate buffer from defatted seed flour of Albizia lebbeck. It was purified to 29.62 folds with 51.43... A Bowman-Birk inhibitor with activity against gut proteases of Helicoverpa armigera was extracted in 0.I M sodium phosphate buffer from defatted seed flour of Albizia lebbeck. It was purified to 29.62 folds with 51.43% recovery using ammonium sulfate precipitation, gel filtration chromatography on Sephadex G-100 column and ion ex- change chromatography on DEAE-Sephadex As0. The purified protein had a molecular weight of 12,303 daltons as determined by SDS-PAGE. It was found to be heat stable up to 60~C and had two pH optima of 7.5 and 9.0. The inhibitor exhibited non-competitive pattern of inhibition with a low Ki value of 0.2 ~tM. The inhibitoi- was found to be susceptible to varying concentrations of reducing agents like DTT and 2- mercaptoethanol, thereby indicating the role of disulphide bridges in maintaining its three dimensional structure and stability. The purified inhibitor caused mortality and suppressed larval growth ofPieris brassi- cae larvae. It was also found to be effective against gut trypsin extracted from Spodoptera littoralis. The sequence of the genes encoding for such inhibitors can be determined and the genes expressing protease inhibitors can be used in vegetable crops to confer resistance against insect pests and other plant pathogens. 展开更多
关键词 Albizia lebbeck trypsin inhibitor insecticidal activity purifi-cation Pieris brassicae
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Purification and Trypsin Inhibitor Activity of a Sporamin B from Sweet Potato (Ipomoea batatas Lam. 55-2) 被引量:1
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作者 SUN Yan-li SUN Jun-mao LI Qing-peng 《Agricultural Sciences in China》 CAS CSCD 2009年第7期808-820,共13页
Sporamin is a soluble protein in sweet potato, and falls into two distinct homology groups, subfamilies A and B. In this research, a sporamin B was purified and its amino acid sequences, trypsin inhibitor activity (T... Sporamin is a soluble protein in sweet potato, and falls into two distinct homology groups, subfamilies A and B. In this research, a sporamin B was purified and its amino acid sequences, trypsin inhibitor activity (Ti activity) were analyzed. This sporamin B was isolated from sweet potato tubers [Ipomoea batatas (L.) Lam cv. 55-2] through extraction of the water-soluble fraction, dialysis, ultrafiltration and ion-exchange chromatography. Homology determined by polyacrylamide gel electrophoresis showed that mainly one bond appeared in gel after being reduced by SDS (sodium dodecyl sulfate), or by SDS and 2-mercaptoethanol, or in native situation. By comparing the data of the polypeptide mass Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry with those of the mass of the theoretical amino acid sequences from NCBI protein database, it was revealed that it was Q40091|Q40091_IPOBA, sweet potato sporamin B - Ipomoea Batatas (sweet potato) (Batate). The sequence coverage was 70.6%. N-terminal sequence was SETPV (Ser-Glu-Thr-Pro-Val). There is a linear relationship between trypsin inhibitor activity (Ti activity) and amounts of this sporamin B (3-18 μg mL-1). The equation of linear regression was y = 2.5809x + 17.049 (r2 = 0.9966). There was a curvilinear relationship between Ti activity and amounts of this sporamin B (21-150 μg mL-1). The equation of curvilinear regression is y = 14.417ln(x) + 23.26 (r2 = 0.9924). The concentration of sporamin B with Ti activity after heating at 40°C may induce part denature of this sporamin B, and there was no statistic difference after heating at 40, 50, 60°C for 20 min. Heat treatment at more than 90°C leads to a dramatic decrease of trypsin inhibitor efficiency. The results suggested that Q40091|Q40091_IPOBA was the major sporamin B in sweet potato tubers [Ipomoea Batatas (L.) Lam cv. 55-2], which had strong Ti activity, and was stable to both thermal and DTT (DL-dithiothreitol) relatively. 展开更多
关键词 sweet potato sporamin B IDENTIFICATION trypsin inhibitor activity
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Pancreatic secretory trypsin inhibitor:More than a trypsin inhibitor 被引量:2
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作者 Gai-Ping Wang,Cun-Shuan Xu,College of Life Science,Henan Normal University,Xinxiang 453007,Henan Province,China Co-construction Key Laboratory for Cell Differentiation and Regulation,Henan Normal University,Xinxiang 453007,Henan Province,China 《World Journal of Gastrointestinal Pathophysiology》 CAS 2010年第2期85-90,共6页
Kazal-type serine protease inhibitor is one of the most important and widely distributed protease inhibitor families. Pancreatic secretory trypsin inhibitor (PSTI), also known as serine protease inhibitor Kazal type I... Kazal-type serine protease inhibitor is one of the most important and widely distributed protease inhibitor families. Pancreatic secretory trypsin inhibitor (PSTI), also known as serine protease inhibitor Kazal type I(SPINK1), binds rapidly to trypsin, inhibits its activity and is likely to protect the pancreas from prematurely activated trypsinogen. Therefore, it is an important factor in the onset of pancreatitis. Recent studies found that PSTI/SPINK1 is also involved in self-regulation of acinar cell phagocytosis, proliferation and growth of a variety of cell lines. In addition, it takes part in the response to inflammatory factor or injury and is highly related to adult type II citrullinemia. 展开更多
关键词 Pancreatic secretory trypsin inhibitor/serine protease inhibitor Kazal type I PANCREATITIS AUTOPHAGY Cell proliferation Inflammatory factor Adult-II citrullinemia.
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Relationship between inter-α-trypsin inhibitor heavy chain 4 and ovarian cancer 被引量:2
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作者 Min Huang Wei Zhang +1 位作者 Bingbing Zhao Li Li 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2019年第6期955-964,共10页
Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study a... Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study aimed to explore the effect of ITIH4 on OC and to identify its underlying mechanism.Methods:Expressions of ITIH4 in OC tissues and cells were determined using quantitative reverse transcription polymerase chain reaction(RT-qPCR)and western blots.The function of ITIH4 in the OC cell line HO8910 pm was tested via ITIH4 knockdown.The cell growth rate was measured using MTT and colony formation assays.Flow cytometry was performed to evaluate cell cycle progression.Cell migration and invasion abilities were observed using the transwell migration assay.Results:ITIH4 was downregulated in OC tissues and cells.ITIH4 knockdown promoted cell growth and cell cycle progression.Consistent with these results,inhibition of ITIH4 in OC cells significantly increased cell migration and invasion abilities.Cox regression analysis suggests that ITIH4 expression alone is not a good predictor of the prognosis of malignant ovarian tumors in patients.Conclusions:ITIH4 inhibits the progression of OC,suggesting that ITIH4 may be a useful biomarker for OC.This study may provide a potential novel target for the treatment of OC. 展开更多
关键词 Ovarian cancer inter-α-trypsin inhibitor H4 protein quantitative reverse transcription PCR RNAI
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Silica-supported Macroporous Chitosan Bead for Affinity Purification of Trypsin Inhibitor
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作者 Feng Na XI Jian Min WU Ming Ming LUAN 《Chinese Chemical Letters》 SCIE CAS CSCD 2005年第8期1089-1092,共4页
Macroporous cross-linking chitosan layer coated on silica gel (CTS-SiO2) was prepared by phase inversion and polyethylene glycol (PEG)molecular imprinting methods. Formation of macroporous surtace was investigated... Macroporous cross-linking chitosan layer coated on silica gel (CTS-SiO2) was prepared by phase inversion and polyethylene glycol (PEG)molecular imprinting methods. Formation of macroporous surtace was investigated by scanning electron microscopy (SEM) and BET analysis.The prepared bead was activated by reacting with 1,2-ethylene digiycidyl ether for introducing epoxy groups, and trypsin could be efficiently immobilized on the bead as a biospecific ligand.The bead bearing trypsin was employed to purify trypsin inhibitor (TIs) from egg white as affinity adsorbent. 展开更多
关键词 CHITOSAN SILICA trypsin inhibitor trypsin affinity purification.
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A Novel Trypsin Inhibitor-Like Cysteine-Rich Peptide from the Frog Lepidobatrachus laevis Containing ProteinaseInhibiting Activity
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作者 Yu-Wei Wang Ji-Min Tan +4 位作者 Can-Wei Du Ning Luan Xiu-Wen Yan Ren Lai Qiu-Min Lu 《Natural Products and Bioprospecting》 CAS 2015年第4期209-214,共6页
Various bio-active substances in amphibian skins play important roles in survival of the amphibians.Many protease inhibitor peptides have been identified from amphibian skins,which are supposed to negatively modulate ... Various bio-active substances in amphibian skins play important roles in survival of the amphibians.Many protease inhibitor peptides have been identified from amphibian skins,which are supposed to negatively modulate the activity of proteases to avoid premature degradation or release of skin peptides,or to inhibit extracellular proteases produced by invading bacteria.However,there is no information on the proteinase inhibitors from the frog Lepidobatrachus laevis which is unique in South America.In this work,a cDNA encoding a novel trypsin inhibitor-like(TIL)cysteine-rich peptide was identified from the skin cDNA library of L.laevis.The 240-bp coding region encodes an 80-amino acid residue precursor protein containing 10 half-cysteines.By sequence comparison and signal peptide prediction,the precursor was predicted to release a 55-amino acid mature peptide with amino acid sequence,IRCPKDKIYKFCGSPCPPSCKDLTPNCIAVCKKGCFCRDGTVDNNHGKCVKKENC.The mature peptide was named LL-TIL.LL-TIL shares significant domain similarity with the peptides from the TIL supper family.Antimicrobial and trypsin-inhibitory abilities of recombinant LL-TIL were tested.Recombinant LL-TIL showed no antimicrobial activity,while it had trypsin-inhibiting activity with aKi of 16.5178 lM.These results suggested there was TIL peptide with proteinase-inhibiting activity in the skin of frog L.laevis.To the best of our knowledge,this is the first report of TIL peptide from frog skin. 展开更多
关键词 trypsin inhibitor Cysteine-rich peptide AMPHIBIAN Skin
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CRYSTAL STRUCTURE OF THE COMPLEX OF MUNG BEAN TRYPSIN INHIBITOR LYSINE ACTIVE FRAGMENT WITH BOVINE TRYPSIN AT 1.8 A RESOLUTION
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作者 You Qi TANG Gen Pei LI Zhong Guo CHEN Jie ZENG(Institute of Physical Chemistry,Peking University,Beijing 100871)Tien Chin TSAO Guang Da LIN Rong Guang ZHANG Zheng Wu CHI(Institute of Biochemistry,Academic Sinica,Shanghai 200031) 《Chinese Chemical Letters》 SCIE CAS CSCD 1990年第1期61-64,共4页
The structure of the complex of mung bean trypsin inhibitor lysine active fragment with bovine trypsin has been determined at a resolution of 1.8 A by A-ray crystallographic analysis and the complex model refined by r... The structure of the complex of mung bean trypsin inhibitor lysine active fragment with bovine trypsin has been determined at a resolution of 1.8 A by A-ray crystallographic analysis and the complex model refined by restrained least-squares minimization with the data between 10 and 1.8 resolution.The current conventional R factor is 17.3%,and the model con- tains 1648 protein atoms,219 inhibitor atoms and 126 water molecules.The most prominent feature of the inhibitor fragment is that it does not contain any alpha-helices.Most of the chain fold in an irregular fashion.The seven residues of the binding segment of the inhibitor lysine active frag- ment are in specific contact with bovine trypsin.The binding interaction and geometry around the reactive site are similar to that observed in other studies of trypsin-inhibitor complexes. 展开更多
关键词 maps CRYSTAL STRUCTURE OF THE COMPLEX OF MUNG BEAN trypsin inhibitor LYSINE ACTIVE FRAGMENT WITH BOVINE trypsin AT 1.8 A RESOLUTION AT
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Storage Proteins and Trypsin Inhibitors of an Underutilized Legume, <i>Mucuna</i>: Variability and Their Stability during Germination 被引量:1
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作者 Kempohalli S. Chandrashekharaiah 《American Journal of Plant Sciences》 2013年第4期910-916,共7页
The proteins and trypsin inhibitors were isolated from the seeds of different varieties/accessions of an underutilized legume, Mucuna. The crude protein content of all the germplasms of Mucuna is varied from 15% - 26%... The proteins and trypsin inhibitors were isolated from the seeds of different varieties/accessions of an underutilized legume, Mucuna. The crude protein content of all the germplasms of Mucuna is varied from 15% - 26%, showed little variation and contain higher crude protein when compared with other Mucuna species reported earlier and the pulse crops commonly consumed in India. The seeds of all the varieties of Mucuna exhibited trypsin inhibitor activity. The trypsin inhibitor activity varied from 11 - 14 TIA/mg of protein. Not much variation was observed in trypsin inhibitory activities in soaked seeds compared to dry seeds. Germination of Mucuna pruriens has been carried out and the change in the protein content and trypsin inhibitors were monitored. The protein content of the endosperm increased up to 72 hrs of germination and then decreased. The trypsin inhibitory activity decreased with increase in germination time. The trypsin inhibitor activity was decreased from 14.81 TIA/mg to 2.62 TIA/mg (82% reduction in the trypsin inhibitor activity) after 144 hrs germination. 展开更多
关键词 MUCUNA Seeds Proteins trypsin inhibitors GERMINATION Variation
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A Kunitz trypsin inhibitor from chickpea (<i>Cicer arietinum</i>L.) that exerts an antimicrobial effect on Fusarium oxysporum f.sp. ciceris
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作者 Meera Nair Sardul Singh Sandhu 《Agricultural Sciences》 2013年第11期585-594,共10页
Fusarium oxysporum f.sp. ciceris (Foc) is one of the most important fungal pathogens of chickpea and is regarded as a constant threat in tropical and subtropical countries. In order to correlate Fusarium wilt resistan... Fusarium oxysporum f.sp. ciceris (Foc) is one of the most important fungal pathogens of chickpea and is regarded as a constant threat in tropical and subtropical countries. In order to correlate Fusarium wilt resistance/susceptibility in Cicer arietinum to the presence or absence of trypsin inhibitor (TI) in the crude extract, trypsin inhibitory assay (TIA) and in vitro activity of TI against Foc were studied. In the present study, a 20 kDa trypsin inhibitor was purified from Fusarium wilt resistant cultivar (viz. JG 2001-12) by ammonium sulfate precipitation, dialysis and chromatographies with Sephadex G-100 and Diethyl aminoethyl cellulose (DEAE-cellulose-52) ion-exchange column. Results of pathogenecity assay were found to be in correlation to the trypsin inhibitor assay where the Fusarium wilt resistant cultivar showed high trypsin inhibitory activity (99%) in the presence of trypsin enzyme using both natural and synthetic substrates. Preliminary studies using crude extracts of JG 2001-12 showed a decrease in radial growth of Foc. A 45%-82% reduction in conidium germination at 20 μg&middotmL-1?Cicer arietinum trypsin inhibitor (CaTI) concentration was observed, thereby, indicating the use of CaTI in suppression of pathogen and in its deployment through transgenic plants for the management of Fusarium wilt. 展开更多
关键词 CICER arietinum Fusarium Wilt KUNITZ PROTEINASE inhibitor trypsin inhibitor
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An HPLC method for the assay of trypsin inhibitors and its application to the study of Momordica cochinchinensis extract 被引量:1
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作者 林志燕 余祝宏 +3 位作者 陈晨 董琦鑫 王銘章 郁韵秋 《Journal of Chinese Pharmaceutical Sciences》 CAS 2011年第4期397-403,共7页
A reliable and validated HPLC method was established for the assay of trypsin inhibitors(TI)and it was used in the investigation of the active TI components inMomordica cochinchinensis(Cucurbitaceae family).The un... A reliable and validated HPLC method was established for the assay of trypsin inhibitors(TI)and it was used in the investigation of the active TI components inMomordica cochinchinensis(Cucurbitaceae family).The underlying principle of the assay is the measurement of the decrease in trypsin activity due to the presence of TI in analyzed samples,which was achieved by using HPLC separation and quantification of p-nitroanilide that was generated by tryptic hydrolysis of N-α-benzoyl-DL-arginine -4-nitroanilide.The results showed that the HPLC method had higher selectivity than conventional spectrophotometric assay. 展开更多
关键词 HPLC Momordica cochinchinensis trypsin inhibitor
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Vegetative Storage Protein with Trypsin Inhibitor Activity Occurs in Sapindus mukorassi,a Sapindaceae Deciduous Tree 被引量:2
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作者 Shi-Biao Liu Xu-Chu Wang +3 位作者 Min-Jing Shi Yue-Yi Chen Zheng-Hai Hu Wei-Min Tian 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2009年第4期352-359,共8页
A vegetative storage protein (VSP) with trypsin inhibitor activity in a deciduous tree, Sapindus mukorassi, was characterized by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western-blot, immu... A vegetative storage protein (VSP) with trypsin inhibitor activity in a deciduous tree, Sapindus mukorassi, was characterized by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western-blot, immuno-histochemical localization, light- and electro-microscopy, together with analysis of proteinase inhibitor activity of the purified VSP in vitro. There were two proteins with molecular masses of about 23 and 27 kDa in a relatively high content in the bark tissues of terminal branches of S. mukorassi in leafless periods. The proteins decreased markedly during young shoot development, indicating their role in seasonal nitrogen storage. Immuno-histochemical localization with the polyclonal antibodies raised against the 23 kDa protein demonstrated that the 23 kDa protein was the major component of protein inclusions in protein-storing cells. The protein inclusions were identified by protein-specific staining and should correspond to the electron-dense materials in different forms in the vacuoles of phloem parenchyma cells and phloem ray parenchyma cells under an electron microscope. So, the 23 kDa protein was a typical VSP in S. mukorassi. The 23 and 27 kDa proteins shared no immuno-relatedness, whereas the 23 kDa protein was immuno-related with the 22 kDa VSP in lychee and possessed trypsin inhibitor activity. The 23 kDa protein may confer dual functions: nitrogen storage and defense. 展开更多
关键词 annual growth cycle Chinese soapberry IMMUNO-LOCALIZATION seasonal nitrogen storage trypsin inhibitor.
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TOTAL SYNTHESIS OF Trichosanthes TRYPSIN INHIBITOR AND ITS ANALOGUE 被引量:1
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作者 黄种福 吴明磊 戚正武 《Science China Chemistry》 SCIE EI CAS 1990年第10期1192-1200,共9页
Trichosanthes trypsin inhibitor (TTI) is a peptide consisting of 27 amino acid residues with three pairs of disulfide bonds. This paper reports the total synthesis and disulfide bond refolding of this inhibitor and it... Trichosanthes trypsin inhibitor (TTI) is a peptide consisting of 27 amino acid residues with three pairs of disulfide bonds. This paper reports the total synthesis and disulfide bond refolding of this inhibitor and its analogue. After purification, the amino acid sequence and stoichiometrical inhibitory activity against trypsin of the synthetic inhibitor were compatible with those of the natural inhibitor. The analogue of this inhibitor in which residue Met in position 6 was replaced by Ala was also synthesized. The antitrypsin activity of this synthetic analogue was also approximate to that of the natural inhibitor. 展开更多
关键词 peptide synthesis squash family inhibitors Trichosanthes trypsin inhibitor disulfide bond refolding
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The amino acid sequence of a double-headed trypsin inhibitor from the seeds of Momordica charantia Linn.Cucurbitaceae
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作者 ZENG Fu-Yue QIAN Rui-Qing WANG Yu Shanghai Institute of Organic Chemistry,Academia Sinica.Shanghai 《Acta Chimica Sinica English Edition》 SCIE CAS CSCD 1989年第1期79-85,共9页
A double-headed trypsin inhibitor(MCI-1)was isolated and purified from the seeds of Momordica charantia Linn.Cucurbitaceae,by using the trypsin-sepharose-4B affinity chroma- tography and CM-Sephadex-C50 ion exchange c... A double-headed trypsin inhibitor(MCI-1)was isolated and purified from the seeds of Momordica charantia Linn.Cucurbitaceae,by using the trypsin-sepharose-4B affinity chroma- tography and CM-Sephadex-C50 ion exchange chromatography.It is composed of 77 amino acid residues:Asp_8 Thr_1 Ser_4 Glu_8 Pro_2 Gly_6 Ala_4 Cys_(14) Val_2 Met_4 Ile_8 Leu_1 Phe_1 His_3 Lys_ Arg_7. The amino acid sequence of MCI-1 was determined by sequencing the cyanogen bromide,tryptic and staphylococcus aureus V8 proteolytic peptides,then aligned by overlapped sequences.The result shows that MCI-1 contains 7 pairs of disulfide bonds,its sequence showed the high homology with those of “Bowman-Birk”inhibitors.About 50% trypsin inhibitory activity still remained after MCI-1 was cleavaged with cyanogen bromide. 展开更多
关键词 MCI The amino acid sequence of a double-headed trypsin inhibitor from the seeds of Momordica charantia Linn.Cucurbitaceae ACID
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STUDIES ON NATURAL AND MODIFIED PEPTIDE Trichosanthes TRYPSIN INHIBITORS 被引量:1
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作者 钱岳伟 谭复隆 戚正武 《Science China Chemistry》 SCIE EI CAS 1990年第5期599-605,共7页
A peptide trypsin inhibitor was isolated and purified from the roots of Trichosanthes kirilowii (a Chinese medical herb) by using immobilized anhydro-trypsin affinity chromatography and HPLC C_(18) column reverse chro... A peptide trypsin inhibitor was isolated and purified from the roots of Trichosanthes kirilowii (a Chinese medical herb) by using immobilized anhydro-trypsin affinity chromatography and HPLC C_(18) column reverse chromatography. It contains two major components, both consisting of 27 amino acid residues with three pairs of disulfide bonds. The sequence determination indicated that the difference between them is only in the ninth position, being Gln and Lys, respectively. The peptide bond of the inhibitor reactive site Arg-Ile (3--4) is easy to cleave at low pH by trypsin, resulting in a modified inhibitor. It might be the smallest naturally occurring protein inhibitor so far known. The modification reaction of the Trichosanthes inhibitor with trypsin is similar to the catalytic enzyme-substrate reaction. The dissociation constant of the modified inhibitor with trypsin is around fourfold that of the natural inhibitor. 展开更多
关键词 Trichosanthes trypsin inhibitor ENZYME modification AMINO acid sequence.
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Mung Bean Trypsin Inhibitor——Synthesis of a Fragment and Its Analogues
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作者 李一莉 林晓红 +2 位作者 崔大敷 唐有祺 戚正武 《Science China Chemistry》 SCIE EI CAS 1994年第10期1208-1215,共8页
The Lys active fragment of the mung bean trypsin inhibitor is composed of two peptide chains,one with 26 amino acid residues and the other with 9 residues linked by two interdisulfide bonds. The two peptide chains cou... The Lys active fragment of the mung bean trypsin inhibitor is composed of two peptide chains,one with 26 amino acid residues and the other with 9 residues linked by two interdisulfide bonds. The two peptide chains could be separated successfully from each other by reduction.with DTT followed by gel filtration.The reduced long peptide chain containing 6 Cys residues was subjected to air oxidation, and about 25% of the original antitrypsin activity of the Lys fragment was recovered.Following the previ- ously reported sequence,the solid-phase synthesis of this long peptide chain and its disulfide bond refold- ing are presented.Unexpectedly,the synthetic peptide showed much lower antitrypsin activity than the natural one after reduction and air reoxidation.In order to explain this uncompatible result,we redeter- mined the sequence of the native long peptide chain of the Lys active fragment and obtained the result that the P′_2 position is Ile instead of Lys as previously reported.To ascertain the correct sequence,we synthe- sized another 22-peptide following the newly determined 26-peptide sequence,and skimming two residues respectively from N-terminus and C-terminus.After reduction and reoxidation,the synthetic 22-peptide had the same antitrypsin activity as that of the native 26-peptide.Meanwhile,an analogue of this 22-pep- tide in which the residue Lys at the reactive site was replaced by Ala was also synthesized.This synthetic analogue did not show any activity either to trypsin or to elastase. 展开更多
关键词 peptide synthesis Bowman-Birk inhibitor MUNG BEAN trypsin inhibitor DISULFIDE bond REFOLDING
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血清LncRNA FAF、ITIH4在慢性心力衰竭患者中的表达意义及对预后的预测价值
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作者 江洋 刘泽岩 +3 位作者 叶丽 汪栋林 程景林 万俊 《疑难病杂志》 CAS 2024年第4期418-422,428,共6页
目的探讨血清长链非编码RNA(LncRNA)FAF、间α胰蛋白酶抑制因子重链4(ITIH4)在慢性心力衰竭(CHF)患者中的表达意义及对预后的预测价值。方法选择2020年1月—2022年1月安徽医科大学第二附属医院急诊内科收治的CHF患者187例为CHF组,再根据... 目的探讨血清长链非编码RNA(LncRNA)FAF、间α胰蛋白酶抑制因子重链4(ITIH4)在慢性心力衰竭(CHF)患者中的表达意义及对预后的预测价值。方法选择2020年1月—2022年1月安徽医科大学第二附属医院急诊内科收治的CHF患者187例为CHF组,再根据NYHA心功能分级分为Ⅱ级亚组65例,Ⅲ级亚组77例,Ⅳ级亚组45例。于同期招募健康志愿者103例为健康对照组。2组受试者均检测血清LncRNA FAF表达和ITIH4水平,CHF患者出院后随访12个月,统计随访期间主要不良心血管事件(MACE)发生率;多因素Logistic回归分析影响CHF患者发生MACE的因素;受试者工作特征(ROC)曲线分析LncRNA FAF、ITIH4预测CHF患者发生MACE的价值。结果CHF组血清LncRNA FAF表达、ITIH4水平低于健康对照组(t/P=24.469/<0.001、35.196/<0.001)。血清LncRNA FAF表达、ITIH4水平Ⅳ级亚组低于Ⅲ级亚组低于Ⅱ级亚组(F/P=91.653/<0.001、102.345/<0.001)。MACE亚组血清LncRNA FAF表达,ITIH4水平低于非MACE亚组(t/P=13.556/<0.001、6.293/<0.001)。NYHAⅣ级、高水平NT-proBNP是CHF患者发生MACE的危险因素[OR(95%CI)=4.627(2.245~9.538)、2.284(1.505~3.468)],高表达LncRNA FAF、高水平ITIH4是保护因素[OR(95%CI)=0.599(0.425~0.844)、0.666(0.478~0.928)]。LncRNA FAF、ITIH4及二者联合预测CHF患者发生MACE的曲线下面积为0.796、0.801、0.896,二者联合预测曲线下面积高于单独预测(Z=2.453、2.404,均P<0.001)。结论CHF患者血清LncRNA FAF表达和ITIH4水平均降低,且与不良预后有关,联合LncRNA FAF和ITIH4可预测CHF患者预后不良风险。 展开更多
关键词 慢性心力衰竭 成纤维细胞生长因子相关因子 间α胰蛋白酶抑制因子重链4 预后
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