Disulfide bonds in recombinant human platelet-derived growth factor BB dimer: Characterization of intermolecular and intramolecular disulfide linkages

Interchain cystines of PDGF-BB dimer were characterized by Edman reaction and by SDS-PAGE analysis on the protein which was chemically cleaved at Trp-40. It was found that Cys-43 has a key role in dimer formation, asymmetrically cross-linked to a cysteine residue of another identical subunit. The re...

Full description

Saved in:
Bibliographic Details
Published in:Biochemistry (Easton) 1993-03, Vol.32 (9), p.2431-2437
Main Authors: Haniu, Mitsuru, Rohde, Michael F, Kenney, William C
Format: Article
Language:English
Subjects:
Acetylation
Alkylation
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Becaplermin
Biological and medical sciences
Cysteine - analysis
Disulfides - chemistry
Fundamental and applied biological sciences. Psychology
Humans
Molecular Sequence Data
Oxidation-Reduction
Peptide Fragments - chemistry
Platelet-Derived Growth Factor - chemistry
Protein Conformation
Protein hormones. Growth factors. Cytokines
Proteins
Proto-Oncogene Proteins c-sis
Recombinant Proteins - chemistry
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Interchain cystines of PDGF-BB dimer were characterized by Edman reaction and by SDS-PAGE analysis on the protein which was chemically cleaved at Trp-40. It was found that Cys-43 has a key role in dimer formation, asymmetrically cross-linked to a cysteine residue of another identical subunit. The remaining cystines participate in the intramolecular disulfide linkages. Pepsin digestion of PDGF-BB dimer generated several small peptides and one ubiquitous Cys-containing peptide. Sequence analyses of several Cys-containing peptides indicated the existence of three intramolecular disulfide linkages including Cys-16--Cys-60, Cys-49--Cys-97, and Cys-53--Cys-99. Two interchain disulfide bonds of Cys-43--Cys-52 between two subunits were deduced from the partial reduction and alkylation of PDGF-BB. This study provides chemically determined disulfide linkages of PDGF-BB.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00060a039