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PMID: 7799941 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Effects of phosphorylation by CAK on cyclin binding by CDC2 and CDK2.

Molecular and cellular biology ·Vol. 15 ·No. 1 ·1995-01-00 ·Pages 345-50

Desai D, Wessling HC, Fisher RP, Morgan DO

Abstract

The cyclin-dependent protein kinases (CDKs) are activated by association with cyclins and by phosphorylation at a conserved threonine residue by the CDK-activating kinase (CAK). We have studied the binding of various human CDK and cyclin subunits in vitro, using purified proteins derived from baculovirus-infected insect cells. We find that most CDK-cyclin complexes known to exist in human cells (CDC2-cyclin B, CDK2-cyclin A, and CDK2-cyclin E) form with high affinity in the absence of phosphorylation or other cellular components. One complex (CDC2-cyclin A) forms with high affinity only after CAK-mediated phosphorylation of CDC2 at the activating threonine residue. CDC2 does not bind with high affinity to cyclin E in vitro, even after phosphorylation of the CDC2 subunit. Thus, phosphorylation is of varying importance in the formation of high-affinity CDK-cyclin complexes.

MeSH Terms
CDC2 Protein Kinase/metabolism CDC2-CDC28 Kinases Chromatography, Gel Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases/metabolism Cyclins/metabolism Humans In Vitro Techniques Mutagenesis, Site-Directed Phosphorylation Protamine Kinase/metabolism Protein Serine-Threonine Kinases/metabolism Recombinant Proteins Structure-Activity Relationship
Chemicals
Cyclins Recombinant Proteins Protamine Kinase Protein Serine-Threonine Kinases CDC2 Protein Kinase CDC2-CDC28 Kinases CDK2 protein, human Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases cyclin-dependent kinase-activating kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Desai D
Department of Physiology, University of California, San Francisco 94143-0444.
Wessling H C
Fisher R P
Morgan D O
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.1.345
Published
1995-01-00
Pages
345-50
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231966
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.15.1.345 PMC Full Text View journal details
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