Protein inhibitor of cAMP-dependent protein kinase: production and characterization of antibodies and intracellular localization.

Published

Journal Article

Multiple forms of protein kinase inhibitor exist in mammalian testis. Specific antibodies to testicular protein kinase inhibitor (PKI) have been raised in sheep. The antibody to the smallest of the inhibitors (9300 daltons) has been purified by antigen-affinity chromatography and shown to give a precipitin band with the inhibitor by double immunodiffusion. The antibody does not recognize any of the subunits of cyclic nucleotide-dependent protein kinases, namely cGMP-dependent protein kinase or the catalytic or regulatory subunits from type I or type II cAMP-dependent protein kinases. The biological activity of the 9300 dalton PKI is blocked completely by a 5 fold molar excess of antibody. Furthermore, the antibody can also block the activity of all other forms of testicular PKI. Using the antibody in indirect immunofluorescence microscopy, PKI localization was examined during interphase and mitosis in a variety of cell types. Our observations indicate that PKI is localized on microtubules in the cytoplasmic microtubule complex during interphase and in the spindle apparatus during mitosis. We suggest that PKI may play a role in the cAMP-dependent regulation of microtubule structure and/or function.

Full Text

Duke Authors

Cited Authors

  • Tash, JS; Welsh, MJ; Means, AR

Published Date

  • August 1, 1980

Published In

Volume / Issue

  • 21 / 1

Start / End Page

  • 57 - 65

PubMed ID

  • 6250718

Pubmed Central ID

  • 6250718

International Standard Serial Number (ISSN)

  • 0092-8674

Digital Object Identifier (DOI)

  • 10.1016/0092-8674(80)90114-2

Language

  • eng

Conference Location

  • United States