Structural analysis of tissue inhibitor of metalloproteinases-1 (TIMP-1) by tryptic peptide mapping.
Tryptic digests of recombinant TIMP-1 have been resolved on reverse-phase HPLC and the major peaks identified by N-terminal sequencing. This procedure accounted for the entire molecule, except two short peptides of 2 and 4 amino acids in length. The peptide map was used to (i), characterize an insoluble 'core' peptide seen on digestion of TIMP-1 in non-reducing conditions; (ii), confirm the structure of delta 127-184TIMP-1, a recently described truncated form of the TIMP-1 molecule; (iii), identify exposed regions of the intact and truncated TIMP-1 molecules by measuring the rate of tryptic peptide release and (iv), locate sites of aberrant proteolysis seen when recombinant human TIMP-1 was purified at large scale.
Biological Laboratory, University of Kent, Canterbury, UK.
R A
RA
Williamson
B J
BJ
Smith
S
S
Angal
G
G
Murphy
R B
RB
Freedman
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004220:Disulfides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000737:chemistry; D019714:Tissue Inhibitor of Metalloproteinases; D014357:Trypsin
D004220:Disulfides; D006023:Glycoproteins; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D014357:Trypsin