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Title
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Disulphide bond assignment in human tissue inhibitor of metalloproteinases (TIMP).
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Authors
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R.A.Williamson,
F.A.Marston,
S.Angal,
P.Koklitis,
M.Panico,
H.R.Morris,
A.F.Carne,
B.J.Smith,
T.J.Harris,
R.B.Freedman.
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Ref.
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Biochem J, 1990,
268,
267-274.
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PubMed id
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Abstract
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Disulphide bonds in human recombinant tissue inhibitor of metalloproteinases
(TIMP) were assigned by resolving proteolytic digests of TIMP on reverse-phase
h.p.l.c. and sequencing those peaks judged to contain disulphide bonds by virtue
of a change in retention time on reduction. This procedure allowed the direct
assignment of Cys-145-Cys-166 and the isolation of two other peptides containing
two disulphide bonds each. Further peptide cleavage in conjunction with
fast-atom-bombardment m.s. analysis permitted the assignments Cys-1-Cys-70,
Cys-3-Cys-99, Cys-13-Cys-124 and Cys-127-Cys-174 from these peptides. The sixth
bond Cys-132-Cys-137 was assigned by inference, as the native protein has no
detectable free thiol groups.
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