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PMID: 2175907 Published · ppublish English Journal Article

Identification of mitosis-specific p65 dimer as a component of human M phase-promoting factor.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 24 ·1990-12-00 ·Pages 9600-4

Meikrantz W, Suprynowicz FA, Halleck MS, Schlegel RA

Abstract

Antisera raised against two mitosis-specific protein kinases from human cells recognized a single 65-kDa polypeptide (p65) that is present in similar amounts in interphase and mitotic cell extracts. Immunoblot analysis of reduced and unreduced extracts revealed that p65 exists as a 65-kDa monomer during interphase but forms a 130-kDa disulfide-linked homodimer during mitosis. Several different antibodies recognizing the p34cdc2 protein kinase and cyclin B components of M phase-promoting factor (MPF) coprecipitated p65 from mitotic but not from interphase extracts. In addition, an anti-p65 immunoaffinity column substantially depleted mitotic extracts of histon H1 kinase activity assayed under conditions diagnostic for MPF. These results suggest that active human MPF may be a complex of p34cdc2, cyclin B, and dimeric p65. A sulfhydryl cycle, proposed in the earlier literature on the biochemistry of mitosis, might underlie the dimerization of p65 and formation of active MPF.

MeSH Terms
Blotting, Western Cell Line Chromatography, Affinity Disulfides/metabolism Humans Immune Sera Immunoglobulin G Interphase Macromolecular Substances Mitosis Molecular Weight Protamine Kinase/metabolism Protein Kinases/isolation & purification,metabolism
Chemicals
Disulfides Immune Sera Immunoglobulin G Macromolecular Substances Protein Kinases Protamine Kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Meikrantz W
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Suprynowicz F A
Halleck M S
Schlegel R A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.24.9600
Published
1990-12-00
Pages
9600-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC55220
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.87.24.9600 PMC Full Text View journal details
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