Linked Life Data

1618839

Source:http://linkedlifedata.com/resource/pubmed/id/1618839

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
19
pubmed:dateCreated
1992-8-5
pubmed:databankReference
pubmed:abstractText
The type II cAMP-dependent protein kinase (PKA) is localized to specific subcellular environments through binding of the dimeric regulatory subunit (RII) to anchoring proteins. Subcellular localization is likely to influence which substrates are most accessible to the catalytic subunit upon activation. We have previously shown that the RII-binding domains of four anchoring proteins contain sequences which exhibit a high probability of amphipathic helix formation (Carr, D. W., Stofko-Hahn, R. E., Fraser, I. D. C., Bishop, S. M., Acott, T. E., Brennan, R. G., and Scott J. D. (1991) J. Biol. Chem. 266, 14188-14192). In the present study we describe the cloning of a cDNA which encodes a 1015-amino acid segment of Ht 31. A synthetic peptide (Asp-Leu-Ile-Glu-Glu-Ala-Ala-Ser-Arg-Ile-Val-Asp-Ala-Val-Ile-Glu-Gln-Val -Lys-Ala-Ala-Tyr) representing residues 493-515 encompasses the minimum region of Ht 31 required for RII binding and blocks anchoring protein interaction with RII as detected by band-shift analysis. Structural analysis by circular dichroism suggests that this peptide can adopt an alpha-helical conformation. Both Ht 31 (493-515) peptide and its parent protein bind RII alpha or the type II PKA holoenzyme with high affinity. Equilibrium dialysis was used to calculate dissociation constants of 4.0 and 3.8 nM for Ht 31 peptide interaction with RII alpha and the type II PKA, respectively. A survey of nine different bovine tissues was conducted to identify RII binding proteins. Several bands were detected in each tissues using a 32P-RII overlay method. Addition of 0.4 microM Ht 31 (493-515) peptide to the reaction mixture blocked all RII binding. These data suggest that all anchoring proteins bind RII alpha at the same site as the Ht 31 peptide. The nanomolar affinity constant and the different patterns of RII-anchoring proteins in each tissue suggest that the type II alpha PKA holoenzyme may be specifically targeted to different locations in each type of cell.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
267
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
13376-82
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:1618839-A Kinase Anchor Proteins, pubmed-meshheading:1618839-Adaptor Proteins, Signal Transducing, pubmed-meshheading:1618839-Amino Acid Sequence, pubmed-meshheading:1618839-Autoradiography, pubmed-meshheading:1618839-Base Sequence, pubmed-meshheading:1618839-Carrier Proteins, pubmed-meshheading:1618839-Circular Dichroism, pubmed-meshheading:1618839-Cloning, Molecular, pubmed-meshheading:1618839-DNA, pubmed-meshheading:1618839-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:1618839-Humans, pubmed-meshheading:1618839-Molecular Sequence Data, pubmed-meshheading:1618839-Mutagenesis, Site-Directed, pubmed-meshheading:1618839-Phosphorylation, pubmed-meshheading:1618839-Protein Conformation, pubmed-meshheading:1618839-Protein Kinases, pubmed-meshheading:1618839-Thyroid Gland
pubmed:year
1992
pubmed:articleTitle
Association of the type II cAMP-dependent protein kinase with a human thyroid RII-anchoring protein. Cloning and characterization of the RII-binding domain.
pubmed:affiliation
Vollum Institute for Advanced Biomedical Research, Oregon Health Sciences University, Portland 97201.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.