Document Detail

Disulfide bond assignments and secondary structure analysis of human and murine interleukin 10.
MedLine Citation:
PMID:  8364028     Owner:  NLM     Status:  MEDLINE    
Interleukin 10 (IL-10), which was first discovered by its ability to inhibit the synthesis of various cytokines, most notably gamma interferon, from Th1 helper cells, displays pleiotropic immunoregulatory properties. Human and murine IL-10 have a high amino acid sequence identity (ca. 73%) which includes the conservation of all four cysteine residues in human IL-10 and the first four out of five cysteine residues for murine IL-10. Chemical analysis was used to determine that both recombinant human and recombinant murine IL-10 contain two disulfide bonds. The disulfide pairs for each were determined by mass spectrometric and reversed-phase HPLC analysis of trypsin-derived polypeptide fragments. The disulfide bond assignments for both species were similar in that the first cysteine residue in the sequence paired with the third and the second paired with the fourth. The fifth cysteine in murine IL-10 was determined by chemical modification to be unpaired. Far-UV circular dichroism analysis indicated that the secondary structure of recombinant human and murine IL-10 are composed of ca. 60% alpha-helix. Reduction of the disulfide bonds structurally destabilized the protein and led to a structure containing only 53% alpha-helix. The reduced protein displayed no in vitro biological activity in a mast cell proliferation assay. These studies indicate that IL-10 is a highly alpha-helical protein containing two disulfide bonds, either one or both of which are critical for its structure and function. In addition, these properties suggest that this interesting cytokine may belong to the alpha helical cytokine class of hematopoietic ligands.
W T Windsor; R Syto; A Tsarbopoulos; R Zhang; J Durkin; S Baldwin; S Paliwal; P W Mui; B Pramanik; P P Trotta
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biochemistry     Volume:  32     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  1993 Aug 
Date Detail:
Created Date:  1993-10-07     Completed Date:  1993-10-07     Revised Date:  2005-11-17    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  8807-15     Citation Subset:  IM    
Schering-Plough Research Institute, Kenilworth, New Jersey 07033.
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MeSH Terms
Amino Acid Sequence
CHO Cells
Chromatography, High Pressure Liquid
Circular Dichroism
Disulfides / analysis*
Interleukin-10 / chemistry*
Molecular Sequence Data
Protein Structure, Secondary*
Recombinant Proteins / chemistry
Spectrophotometry, Ultraviolet
Sulfhydryl Compounds / analysis
Reg. No./Substance:
0/Disulfides; 0/Recombinant Proteins; 0/Sulfhydryl Compounds; 130068-27-8/Interleukin-10

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