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PMID: 6330524 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification, purification, and characterization of phosphotyrosine-specific protein phosphatases from cultured chicken embryo fibroblasts.

Molecular and cellular biology ·Vol. 4 ·No. 6 ·1984-06-00 ·Pages 1003-12

Nelson RL, Branton PE

Abstract

Tyrosine phosphorylation catalyzed by a unique class of protein kinases is an important process in both normal cell proliferation and oncogenic transformation. In this study, phosphoprotein phosphatases specific for the dephosphorylation of phosphotyrosine residues were partially purified from secondary chicken embryo fibroblasts, using 32P-labeled immunoglobulin G phosphorylated by pp60src as substrate. Crude cell extracts contained ca. 70% of the activity in the soluble form and ca. 30% associated with a crude membrane fraction. The soluble activity was purified by using DEAE-cellulose and carboxymethyl cellulose column chromatography and gel filtration, and at least three enzyme species of apparent Mr 55,000 (pTPI), 50,000 (pTPII), and 95,000 (pTPIII)--comprising ca. 20, 45, and 35%, respectively, of the total activity--were resolved. All three enzymes possessed somewhat similar properties. They had a pH optimum of about 7.4, they were inhibited by Zn2+, vanadate, ATP, and ADP, and they were unaffected by divalent metal cations, EDTA, and F- under standard assay conditions employing a physiological ionic strength. These properties suggest that they represent a class of enzymes distinct from well-known phosphoseryl-phosphothreonyl-protein phosphatases and that dephosphorylation of phosphotyrosine-containing proteins may be carried out by a unique family of phosphoprotein phosphatases. Transformation by Rous sarcoma virus resulted in a small increase in phosphotyrosyl-protein phosphatase activity.

MeSH Terms
Animals Avian Sarcoma Viruses/genetics Cell Membrane/enzymology Cell Transformation, Neoplastic Cells, Cultured Chick Embryo Fibroblasts/enzymology Kinetics Molecular Weight Phosphoprotein Phosphatases/isolation & purification,metabolism Phosphorus Radioisotopes Protein Tyrosine Phosphatases
Chemicals
Phosphorus Radioisotopes Phosphoprotein Phosphatases Protein Tyrosine Phosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nelson R L
Branton P E
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43 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.4.6.1003-1012.1984
Published
1984-06-00
Pages
1003-12
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC368867
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.4.6.1003-1012.1984 PMC Full Text View journal details
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