High-level expression of a bioengineered, cysteine-free hepatocyte-stimulating factor (interleukin 6)-like protein

R C Jambou; J N Snouwaert; G A Bishop; J R Stebbins; J A Frelinger; D M Fowlkes

doi:10.1073/pnas.85.24.9426

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High-level expression of a bioengineered, cysteine-free hepatocyte-stimulating factor (interleukin 6)-like protein

Journal article

Open access

Peer reviewed

High-level expression of a bioengineered, cysteine-free hepatocyte-stimulating factor (interleukin 6)-like protein

R C Jambou, J N Snouwaert, G A Bishop, J R Stebbins, J A Frelinger and D M Fowlkes

Proceedings of the National Academy of Sciences - PNAS, Vol.85(24), p.9426

12/1988

DOI: 10.1073/pnas.85.24.9426

PMCID: PMC282765

PMID: 3059347

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Abstract

Hepatocyte-stimulating factor, interferon-beta 2, B-cell stimulation factor 2, and hybridoma/plasmacytoma growth factor are identical proteins presently referred to as interleukin 6 (IL-6). Through the use of synthetic oligonucleotide technology, we have constructed a biologically active recombinant IL-6 (rIL-6) gene based on the sequence of a human IL-6 cDNA. The synthetic gene encodes a cysteine-free, bioengineered rIL-6 protein that is expressed at high levels in Escherichia coli as a tripartite fusion protein. Cleavage of the fusion protein with collagenase releases a 23-kDa rIL-6 protein that can be easily purified to homogeneity. We show that the rIL-6 protein displays a range of biological activities similar to those of natural human IL-6, as demonstrated by its ability to (i) protect cells from viral infection, (ii) stimulate the synthesis of fibrinogen in rat FAZA 967 cells, and (iii) induce the terminal differentiation of B cells, resulting in elevated secretion of immunoglobulin.

Amino Acid Sequence

Cysteine

Recombinant Proteins - analysis

Humans

Molecular Sequence Data

Rats

Antiviral Agents

Interleukins - analysis

Interleukin-6

B-Lymphocytes - drug effects

Interleukins - genetics

Animals

Liver - drug effects

Escherichia coli - genetics

Base Sequence

Interleukins - pharmacology

Details

Title: Subtitle: High-level expression of a bioengineered, cysteine-free hepatocyte-stimulating factor (interleukin 6)-like protein
Creators: R C Jambou - Department of Pathology, University of North Carolina, Chapel Hill 27599
J N Snouwaert
G A Bishop
J R Stebbins
J A Frelinger
D M Fowlkes
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.85(24), p.9426
DOI: 10.1073/pnas.85.24.9426
PMID: 3059347
PMCID: PMC282765
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; United States
Grant note: HL36197 / NHLBI NIH HHS HL07255 / NHLBI NIH HHS HL31012 / NHLBI NIH HHS
Language: English
Date published: 12/1988
Academic Unit: Microbiology and Immunology; President
Record Identifier: 9984002379302771

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