Comparative characterization of receptor and non-receptor associated protein tyrosine kinases.
By using poly(Glu: Tyr, 4:1) as an exogenous substrate, the characteristics of insulin receptor associated protein tyrosine kinase (PTK) from rabbit skeletal muscle has been compared with a growth factor-independent non-receptor PTK partially purified from rat lung particulate fraction. The two PTKs phosphorylated poly(Glu: Tyr; 4:1) very effectively with apparent Km values of 0.3 mg/ml for insulin receptor PTK and 0.8 mg/ml for lung PTK. ATP was the preferred phosphoryl donor for both PTKs (Km = 150 microM); however, in the case of lung PTK, GTP was able to partially replace ATP. ATP analogues, AMP-PNP and ATP-gamma-S inhibited the activities of both enzymes. Receptor PTK was more active in the presence of Mn2+ whereas the lung PTK did not discriminate between Mg2+ or Mn2+ for enzyme activity. Para-hydroxymercurobenzoate (pHMB), a SH-group blocking agent, inhibited the activities of both PTKs, suggesting the requirement of SH-groups for enzymatic activities. Both enzymes were inhibited by fluorosulfonylbenzoyl 5'-adenosine (FSBA). NaCl also inhibited both kinases, however, lung PTK was more sensitive to inhibition. In addition, the lung PTK was not retained on a wheat germ agglutinin (WGA)-agarose column, suggesting that the lung enzyme is either not a glycoprotein or that the carbohydrate moieties present, if any, have no affinity for WGA. Furthermore, the lung PTK appears to be immunologically distinct from both insulin receptor and pp60Src, since it was not immunoprecipitated by antibodies to either pp60Src or insulin receptor. These data indicate that only a few but significant differences exist in the characteristics of receptor and non-receptor associated PTKs.
Srivastava
A K
AK
Laboratory of Diabetes and Carbohydrate Metabolism, Clinical Research Institute of Montreal, Canada.
Chiasson
J L
JL
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002147:Calmodulin / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology; D008168:Lung / Q000201:enzymology; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology; D009712:Nucleotides, Cyclic / Q000494:pharmacology; D010455:Peptides / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D011817:Rabbits; D051381:Rats; D011972:Receptor, Insulin / Q000276:immunology / Q000502:physiology*; D012265:Ribonucleotides / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D013379:Substrate Specificity; D014639:Vanadium / Q000494:pharmacology; D020245:p-Chloromercuribenzoic Acid
D002147:Calmodulin; D002413:Cations, Divalent; D002729:Chloromercuribenzoates; D006023:Glycoproteins; D006133:Growth Substances; D009712:Nucleotides, Cyclic; D010455:Peptides; D012265:Ribonucleotides; D014639:Vanadium; D012965:Sodium Chloride; D020245:p-Chloromercuribenzoic Acid; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D000241:Adenosine