Producing human mechano growth factor (MGF) in Escherichia coli

MGF is a product of a unique muscle-specific splice variant of IGF1 gene (insulin-like growth factor). Its peculiar feature is a specific E-peptide, a 16 a.a. strand at the C-terminus. MGF increases cellular proliferation and inhibits terminal differentiation of myoblasts necessary for the secondary...

Full description

Saved in:
Bibliographic Details
Published in:Protein expression and purification 2008-03, Vol.58 (1), p.70-77
Main Authors: Kuznetsova, Tatiana V., Schulga, Alexey A., Wulfson, Andrey N., Keruchenko, Jan S., Ermolyuk, Yaroslav S., Keruchenko, Irina D., Tikhonov, Roman V., Lisitskaya, Ksenia V., Makarov, Andrey A., Chobotova, Katya, Khomenkov, Vasily G., Khotchenkov, Vyacheslav P., Popov, Vladimir O., Kirpichnikov, Mikhail P., Shevelev, Alexei B.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Cell Proliferation
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression
Growth factor
Humans
Insulin-like growth factor
Molecular Sequence Data
Muscles
Myoblast
Myoblasts - cytology
Plasmids
Proliferation test
Protein Folding
Recombinant protein
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Refolding
STAT5 Transcription Factor - biosynthesis
STAT5 Transcription Factor - chemistry
STAT5 Transcription Factor - genetics
STAT5 Transcription Factor - pharmacology
Tumor Suppressor Proteins - biosynthesis
Tumor Suppressor Proteins - chemistry
Tumor Suppressor Proteins - genetics
Tumor Suppressor Proteins - pharmacology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:MGF is a product of a unique muscle-specific splice variant of IGF1 gene (insulin-like growth factor). Its peculiar feature is a specific E-peptide, a 16 a.a. strand at the C-terminus. MGF increases cellular proliferation and inhibits terminal differentiation of myoblasts necessary for the secondary myotube formation. Previous analysis of physiological effects of MGF was performed using indirect methods such as RT-PCR based examination of the transcript contents in normal tissues, adenovirus-mediated DNA delivery and synthetic E-domain administration. Here, we describe isolation and purification of recombinant MGF thus allowing for the first time the possibility of direct examining MGF effects. The recombinant MGF of directly examining—was expressed in Escherichia coli as inclusion bodies (about 100–200 mg/l), purified and refolded. Biological activity of refolded MGF was analyzed in vitro in proliferation assays with normal human myoblasts. As a result of our work, it has become possible to generate a standard MGF control with characterized activity and a ready-to use MGF test-system neither of which have been previously described. Our data open opportunities for the future works on MGF characterization and to the development of a powerful and highly specific therapeutic agent potentially applicable for muscle growth up-regulation, post-trauma muscle repair, age and hereditary myodystrophy mitigation and in sport medicine.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2007.10.023