A SINGLE MUTATION CONVERTS A NOVEL PHOSPHOTYROSINE BINDING DOMAIN INTO A DUAL-SPECIFICITY PHOSPHATASE

被引:80
作者
WISHART, MJ
DENU, JM
WILLIAMS, JA
DIXON, JE
机构
[1] UNIV MICHIGAN,SCH MED,DEPT BIOL CHEM,ANN ARBOR,MI 48109
[2] UNIV MICHIGAN,SCH MED,DEPT PHYSIOL,ANN ARBOR,MI 48109
[3] UNIV MICHIGAN,SCH MED,DEPT INTERNAL MED,ANN ARBOR,MI 48109
关键词
D O I
10.1074/jbc.270.45.26782
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Dual-specificity protein-tyrosine phosphatases (dsPTPases) have been implicated in the inactivation of mitogen-activated protein kinases (MAPKs), We have identified a novel. phosphoserine/thhreonine/tyrosine-binding protein (STYX) that is related in amino acid sequence to dsPTPases, except for the substitution of Gly for Cys in the conserved dsPTPase catalytic loop (HCXYGXXR(S/T)), cDNA subcloning and Northern blot analysis in mouse shows poly(A(+)) hybridization bands of 4.6, 2.4, 1.5, and 1.2 kilobases, with highest abundance in skeletal muscle, testis, and heart, Polymerase chain reaction amplification of reverse-transcribed poly(A(+)) RNA revealed an alternatively spliced form of STYX containing a unique carboxyl terminus, Bacterially expressed STYX. is incapable of hydrolyzing Tyr(P)-containing substrates; however, mutation of Gly(120) to Cys (G120C), which structurally mimics the active site of dsPTPases, confers phosphatase activity to this molecule, STYX-G120C mutant hydrolyzes p-nitrophenyl phosphate and dephosphorylates both Tyr(P) and Thr(P) residues of peptide sequences of MAPK homologues. The kinetic parameters of dephosphorylation are similar to human dsPTPase, Vaccinia H1-related, including inhibition by vanadate, We believe this is the first example of a naturally occurring ''dominant negative'' phosphotyrosine/serine/threonine-binding protein which is structurally related to dsPTPases.
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页码:26782 / 26785
页数:4
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