Document Detail


Preparation and characterization of a novel exendin-4 human serum albumin fusion protein expressed in Pichia pastoris.
MedLine Citation:
PMID:  17994612     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel recombinant exendin-4 human serum albumin fusion protein (rEx-4/HSA) expressed in Pichia pastoris was prepared and characterized. Ex-4 is a 39-amino acid peptide isolated from the salivary gland of the lizard Heloderma suspectum and is thought to be a novel therapeutic agent for type 2 diabetes. But to gain a continued effect, the peptide has to be injected twice a day owing to its short plasma half-life (t(1/2) = 2.4 h). To extend the half-life of Ex-4 molecule in vivo, we designed a genetically engineered Ex-4/HSA fusion protein. Between Ex-4 and HSA, a peptide linker GGGGS was inserted and the fusion protein was expressed in methylotrophic yeast P. pastoris with native HSA secretion signal sequence. The recombinant protein was secreted correctly and was obtained with high purity (typically > 98%) by a three-step purification procedure. cAMP assay demonstrated that the fusion protein had a bioactivity similar to Ex-4 for interaction with GLP-1 receptors in vitro. Results from oral glucose tolerance test indicated that rEx-4/HSA could effectively improve glucose tolerance in diabetic db/db mice. Pharmacokinetics studies in cynomologus monkeys also showed that rEx-4/HSA had a much longer plasma half-life. Therefore, rEx-4/HSA fusion protein could potentially be used as a new recombinant biodrug for type 2 diabetes therapy.
Authors:
Yan-Shan Huang; Zhi Chen; Yi-Qiong Chen; Guo-Chang Ma; Jian-Feng Shan; Wei Liu; Lin-Fu Zhou
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of peptide science : an official publication of the European Peptide Society     Volume:  14     ISSN:  1075-2617     ISO Abbreviation:  J. Pept. Sci.     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-04-23     Completed Date:  2008-08-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9506309     Medline TA:  J Pept Sci     Country:  England    
Other Details:
Languages:  eng     Pagination:  588-95     Citation Subset:  IM    
Affiliation:
Department of Cell Biology, School of Medicine, Zhejiang University, Hangzhou, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Blood Glucose / metabolism
Cell Line
Cricetinae
Cyclic AMP / biosynthesis
DNA Primers / genetics
Female
Glucose Tolerance Test
Humans
Hypoglycemic Agents / isolation & purification,  pharmacokinetics,  pharmacology
Lizards / genetics
Mice
Mice, Inbred C57BL
Peptides / genetics*,  isolation & purification*,  pharmacokinetics,  pharmacology
Pichia / genetics
Recombinant Fusion Proteins / genetics,  isolation & purification,  pharmacokinetics,  pharmacology
Serum Albumin / genetics*,  isolation & purification*,  pharmacokinetics,  pharmacology
Transfection
Venoms / genetics*,  isolation & purification*,  pharmacokinetics,  pharmacology
Chemical
Reg. No./Substance:
0/Blood Glucose; 0/DNA Primers; 0/Hypoglycemic Agents; 0/Peptides; 0/Recombinant Fusion Proteins; 0/Serum Albumin; 0/Venoms; 141732-76-5/exenatide; 60-92-4/Cyclic AMP

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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