PURIFICATION AND CHARACTERIZATION OF A SOLUBLE CATALYTIC FRAGMENT OF THE HUMAN TRANSMEMBRANE LEUKOCYTE ANTIGEN RELATED (LAR) PROTEIN TYROSINE PHOSPHATASE FROM AN ESCHERICHIA-COLI EXPRESSION SYSTEM

被引:76
作者
CHO, HJ
RAMER, SE
ITOH, M
WINKLER, DG
KITAS, E
BANNWARTH, W
BURN, P
SAITO, H
WALSH, CT
机构
[1] HARVARD UNIV,SCH MED,DEPT BIOL CHEM & MOLEC PHARMACOL,BOSTON,MA 02115
[2] HARVARD UNIV,SCH MED,DANA FARBER CANC INST,BOSTON,MA 02115
[3] F HOFFMANN LA ROCHE & CO LTD,CENT RES DEPT,CH-4002 BASEL,SWITZERLAND
关键词
D O I
10.1021/bi00239a019
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
A 350 amino acid soluble fragment of the intracellular catalytic domain of the human transmembrane leukocyte antigen related (LAR) protein tyrosine phosphatase has been purified 17-fold to > 90% purity from an Escherichia coli expression vector in quantities sufficient for kinetic and structural characterization. To assess substrate specificity, phosphotyrosine peptides corresponding to autophosphorylation sites of the two major classes of tyrosine kinases have been synthesized. Thus 6-12-residue phosphotyrosine peptides of the insulin receptor and epidermal growth factor receptor kinase domains and of the autophosphorylation and C-terminal regulatory sites of p60src and p56lck have been analyzed for k(cat) and K(M) by using a nonradioactive chromogenic assay for P(i) release. The catalytic domain of LAR PTPase shows k(cat) values of 20-70 s-1 for phosphotyrosine peptides and affinities that vary 150-fold from 27-mu-M to 4.1 mM.
引用
收藏
页码:6210 / 6216
页数:7
相关论文
共 47 条
  • [1] A SIMPLE AND EFFECTIVE CHEMICAL PHOSPHORYLATION PROCEDURE FOR BIOMOLECULES
    BANNWARTH, W
    TRZECIAK, A
    [J]. HELVETICA CHIMICA ACTA, 1987, 70 (01) : 175 - 186
  • [2] BERTICS PJ, 1988, J BIOL CHEM, V263, P3610
  • [3] A FUNCTION FOR THE LCK PROTO-ONCOGENE
    BOLEN, JB
    VEILLETTE, A
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1989, 14 (10) : 404 - 407
  • [4] HUMAN-PLACENTA PROTEIN-TYROSINE-PHOSPHATASE - AMINO-ACID SEQUENCE AND RELATIONSHIP TO A FAMILY OF RECEPTOR-LIKE PROTEINS
    CHARBONNEAU, H
    TONKS, NK
    KUMAR, S
    DILTZ, CD
    HARRYLOCK, M
    COOL, DE
    KREBS, EG
    FISCHER, EH
    WALSH, KA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (14) : 5252 - 5256
  • [5] THE LEUKOCYTE COMMON ANTIGEN (CD45) - A PUTATIVE RECEPTOR-LINKED PROTEIN TYROSINE PHOSPHATASE
    CHARBONNEAU, H
    TONKS, NK
    WALSH, KA
    FISCHER, EH
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (19) : 7182 - 7186
  • [6] CDNA ISOLATED FROM A HUMAN T-CELL LIBRARY ENCODES A MEMBER OF THE PROTEIN-TYROSINE-PHOSPHATASE FAMILY
    COOL, DE
    TONKS, NK
    CHARBONNEAU, H
    WALSH, KA
    FISCHER, EH
    KREBS, EG
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (14) : 5257 - 5261
  • [7] AUTOPHOSPHORYLATION SITES ON THE EPIDERMAL GROWTH-FACTOR RECEPTOR
    DOWNWARD, J
    PARKER, P
    WATERFIELD, MD
    [J]. NATURE, 1984, 311 (5985) : 483 - 485
  • [8] HUMAN CDC2 PROTEIN-KINASE IS A MAJOR CELL-CYCLE REGULATED TYROSINE KINASE SUBSTRATE
    DRAETTA, G
    PIWNICAWORMS, H
    MORRISON, D
    DRUKER, B
    ROBERTS, T
    BEACH, D
    [J]. NATURE, 1988, 336 (6201) : 738 - 744
  • [9] PEPTIDE-SYNTHESIS .8. A SYSTEM FOR SOLID-PHASE SYNTHESIS UNDER LOW-PRESSURE CONTINUOUS-FLOW CONDITIONS
    DRYLAND, A
    SHEPPARD, RC
    [J]. JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1, 1986, (01): : 125 - 137
  • [10] PHOSPHORYLATION OF GAP AND GAP-ASSOCIATED PROTEINS BY TRANSFORMING AND MITOGENIC TYROSINE KINASES
    ELLIS, C
    MORAN, M
    MCCORMICK, F
    PAWSON, T
    [J]. NATURE, 1990, 343 (6256) : 377 - 381