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carboxypeptidase A6 Promotes the Proliferation and Migration of Hepatocellular Carcinoma by Up-regulating AKT Signaling Pathway

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Current Medical Science 2019年第5期39卷 727-733页

作者： Qi-bo HUANG Hong-wei ZHANG Zhi-bin LIAO Department of Clinical Medicine Medical College of Wuhan University of Science and TechnologyWuhan 430081China Hepatic Surgery Center Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

Summary:Hepatocellular carcinoma(HCC)has a poor treatment prognosis and high mortality *** the molecular mechanism underlying HCC development would benefit the identification of diagnostic biomarkers and the improvement of the treatment *** expression of carboxypeptidase A6(CPA6)has been reported in epilepsy and febrile seizures rather than in any type of ***,the function of CPA6 expression in HCC is not yet *** this study,we aimed to investigate the clinicopathological significance of the expression of CPA6 in HCC and the underlying *** observed that the expression of the CPA6 protein was increased significantly in HCC tissues than in paracancerous *** explore its function in HCC,both gain-and loss-of-function studies demonstrated that CPA6 played a vital role in promoting HCC growth and *** knocking down CPA6 with shRNA,HCC cell proliferation and migration could be ***,CPA6 overexpression could promote proliferation and migration of HLF ***,CPA6 could activate AKT serine/threonine kinase(AKT)signaling pathway as confirmed by Western *** conclusion,our study revealed that CPA6 could promote HCC cell proliferation and migration via AKT-mediated signaling *** findings suggest that CPA6 is a promising diagnostic biomarker and therapeutic target to improve the prognosis of HCC.

关键词： carboxypeptidase A6 hepatocellular carcinoma proliferation migration AKT

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Comparison of Substrate Specificity of <em>Escherichia Coli</em>p-Aminobenzoyl-Glutamate Hy-drolase with <em>Pseudomonas</em>carboxypeptidase G

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Advances in Enzyme Research 2014年第1期2卷 39-48页

作者： Cassandra M. Larimer Dejan Slavnic Lenore D. Pitstick Jacalyn M. Green Department of Biochemistry Chicago College of Osteopathic Medicine Midwestern University Downers Grove IL USA

Reduced folic acid derivatives support biosynthesis of DNA, RNA and amino acids in bacteria as well as in eukaryotes, including humans. While the genes and steps for bacterial folic acid synthesis are known, those associated with folic acid catabolism are not well understood. A folate catabolite found in both humans and bacteria is p-aminobenzoyl-glutamate (PABA-GLU). The enzyme p-aminobenzoyl-glutamate hydrolase (PGH) breaks down PABA-GLU and is part of an apparent operon, the abg region, in E. coli. The subunits of PGH possess sequence and catalytic similarities to carboxypeptidase enzymes from Pseudomonas species. A comparison of the subunit sequences and activity of PGH, relative to carboxypeptidase enzymes, may lead to a better understanding of bacterial physiology and pathway evolution. We first compared the amino acid sequences of AbgA, AbgB and carboxypeptidase G2 from Pseudomonas sp. RS-16, which has been crystallized. Then we compared the enzyme activities of E. coli PGH and commercially available Pseudomonas carboxypeptidase G using spectrophotometric assays measuring cleavage of PABA-GLU, folate, aminopterin, methotrexate, 5-formyltetrahydrofolate, and 5-methyltetrahydrofolate. The Km and Vmax values for the folate and anti-folate substrates of PGH could not be determined, because the instrument reached its limit before the enzyme was saturated. Therefore, activity of PGH was compared to the activity of CPG, or normalized to PABA-GLU (nmole/min/μg). Relative to its activity with 10 μM PABA-GLU (100%), PGH cleaved glutamate from methotrexate (48%), aminopterin (45%) and folate (9%). Reduced folates leucovorin (5-formyltetrahydrofolate) and 5-methyltetrahydrofolate were not cleaved by PGH. Our data suggest that E. coli PGH is specific for PABA-GLU as its activity with natural folates (folate, 5-methyltetrahydrofolate, and leucovorin) was very poor. It does, however, have some ability to cleave anti-folates which may have clinical applications in treatment

关键词： Folate p-Aminobenzoyl-Glutamate Hydrolase carboxypeptidase G Folate Catabolism

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Solid-Phase Enzymatic Peptide Synthesis to Produce an Antioxidant Dipeptide

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Transactions of Tianjin University 2019年第3期25卷 276-282页

作者： Yuyao Shan Mengfan Wang Wei Qi Rongxin Su Zhimin He State Key Laboratory of Chemical Engineering School of Chemical Engineering and Technology Tianjin University Tianjin Key Laboratory of Membrane Science and Desalination Technology Collaborative Innovation Center of Chemical Science and Engineering

Peptide bond synthesis is favorable to the production of bioactive small peptides. However, the abuse of toxic reagents remains an issue for chemical synthesis method, whereas the low product yield and purity limit the widespread use of enzymatic method. In this study, a new solid-phase enzymatic peptide synthesis(SPEPS) strategy was developed to produce an antioxidant tyrosine-alanine dipeptide(Tyr-Ala) by using recombinant carboxypeptidase Y(CPY) as the catalyst. The general SPEPS procedure involves three steps. First, the N-protected acyl donor was covalently attached to solid resin. Second,the peptide bond was condensed between the acyl donor and the nucleophile under the catalysis of CPY. Finally, one-step cleavage was performed to remove the protecting group and cleave the peptides from solid resin. Upon the optimization of reaction conditions, 77.92%(±2.723%) yield of Tyr-Ala with high product purity of 90.971%(±2.695%) was *** addition, the antioxidant activity of Tyr-Ala was determined by ABTS method, indicating that the synthesized Tyr-Ala obtained by SPEPS showed a superior antioxidant capability compared with commercial glutathione.

关键词： Solid-phase enzymatic peptide synthesis carboxypeptidase Y Tyrosine-alanine Antioxidant activity

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Pancreatic biomarkers:role in diabetes mellitus

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Journal of Pancreatology 2023年第4期6卷 169-177页

作者： Sana Rafaqat Ramsha Hafeez Rida Mairaj Abeerah Saleem Saira Rafaqat Department of Biotechnology(Molecular Human Genetics) Lahore College for Women UniversityLahore 54000Pakistan Department of Zoology(Fisheries and Aquaculture) Lahore College for Women UniversityLahore 54000Pakistan Department of Zoology(Fisheries and Nanotechnology) Lahore College for Women UniversityLahore 54000Pakistan Department of Zoology(Molecular Physiology) Lahore College for Women University54000LahorePakistan

Diabetes mellitus refers to a group of diseases that cause high blood sugar *** most common type is type 2 diabetes,which is caused by insulin resistance and inadequate insulin ***,diabetes can also result from conditions affecting the exocrine *** type 1 and type 2 diabetes patients may experience changes in their pancreatic exocrine function,leading to reduced levels of fecal elastase-1 in many *** review article focuses on the role of specific pancreatic biomarkers in diabetes mellitus,including cholecystokinin,trypsin,chymotrypsin,carboxypeptidase,amylase,lipase,secretin,elastase-1,and retinol-binding protein 4 about recent advances and discoveries,significant gaps in the literature,current debates,and potential directions for future research related to these biomarkers about diabetes *** review article discusses various biomarkers related to pancreatic exocrine and endocrine function and their implications in *** suggests that gut cholecystokinin may play a role in lowering glucose synthesis through a neural network and resistance to it could contribute to hyperglycemia in diabetic *** also discusses the use of various markers such as serum trypsin concentration,amylase and lipase levels,pancreatic elastase levels,and fasting secretin levels to assess pancreatic exocrine ***,the article explores the role of carboxypeptidase E in the endocrine and neurological systems and its association with ***,it also highlights the involvement of retinol-binding protein 4 in the development of type 2 diabetes and insulin resistance.

关键词： Amylase carboxypeptidase Cholecystokinin Chymotrypsin Diabetes Elastase-1 Lipase Pancreatic biomarkers Retinol-binding protein 4 Secretin Trypsin

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Early dynamic transcriptomic changes during preoperative radiotherapy in patients with rectal cancer: A feasibility study

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World Journal of Gastroenterology 2013年第21期19卷 3249-3254页

作者： Stephane Supiot Wilfried Gouraud Loc Campion Pascal Jezéquel Bruno Buecher Josiane Charrier Marie-Francoise Heymann Marc-Andre Mahé Emmanuel Rio Michel Chérel Department of Radiation Oncology Institut de Cancérologie de l'Ouest René Gauducheau44800 Nantes-St-HerblainFrance INSERM U892 Centre de Recherche en Cancérologie Nantes-AngersUniversity of Nantes44000 NantesFrance Centre Hospitalier Universitaire 44000 NantesFrance Department of Nuclear Medicine Institut de Cancérologie de l'Ouest René Gauducheau44800 Nantes-Saint HerblainFrance

AIM: To develop novel biomarkers of rectal radiotherapy, we measured gene expression profiles on biopsies taken before and during preoperative radiotherapy. METHODS: Six patients presenting with a locally advanced rectal cancer (T>T2, N0/Nx, M0) eligible for preoperative radiotherapy (45 Gy in 25 fractions) were selected in a pilot study. Six tumor and 3 normal tissues biopsies were taken before and during radiotherapy,after a dose of 7.2 Gy at a median time of 1 h following irradiation (0:27-2:12). Tumor or normal tissue purity was assessed by a pathologist prior to RNA extraction. Mean RNA content was 23 μg/biopsy (14-37) before radiotherapy and 22.7 μg/biopsy (12-35) during radiotherapy. After RNA amplification, biopsies were analysed with 54K HG-U133A Plus 2.0 Affymetrix expression micro-arrays. Data were normalized according to MAS5 algorithm. A gene expression ratio was calculated as: (gene expression during radiotherapy-gene expression before radiotherapy)/gene expression before radiotherapy. Were selected genes that showed a ratio higher than ± 0.5 in all 6 patients. RESULTS: Microarray analysis showed that preoperative radiotherapy significantly up-regulated 31 genes and down-regulated 6 genes. According to the Gene Ontology project classification, these genes are involved in protein metabolism (ADAMDEC1 ; AKAP7 ; CAPN5 ; CLIC5 ; CPE ; CREB3L1 ; NEDD4L ; RAB27A), ion transport (AKAP7 ; ATP2A3 ; CCL28 ; CLIC5 ; F2RL2 ; NEDD4L ; SLC6A8), transcription (AKAP7 ; CREB3L1 ; ISX ; PAB-PC1L ; TXNIP), signal transduction (CAPN5 ; F2RL2 ; RA- B27A ; TNFRSF11A), cell adhesion (ADAMDEC1 ; PXDN ; SPON1 ; S100A2), immune response (CCL28 ; PXDN ; TNFRSF11A) and apoptosis (ITM2C ; PDCD4 ; PVT1). Up-regulation of 3 genes (CCL28 ; CLIC5 ; PDCD4) was detected by 2 different probes and up-regulation of 2 genes (RAB27A ; TXNIP) by 3 probes. CONCLUSION: Micro-arrays can efficiently assess early transcriptomic changes during preoperative radiotherapy for rectal cancer, and may he

关键词： CCL28 CLIC5 PDCD4 RAB27A TXNIP Protein metabolism Cell adhesion Cell migration SPON1 carboxypeptidase E

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