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Type I cAMP-dependent protein kinase: nucleotide influences upon subunit interactions

Thesis/Dissertation ·
OSTI ID:6985782
Nucleotide binding to both the Type I holoenzyme and the individually isolated subunits was investigated in order to determine the function of these nucleotides in the interactions between the regulatory and catalytic subunits of Type I cAMP-dependent protein kinase. The first part of the study was directed at those interactions in the tetrameric holoenzyme. (..cap alpha../sup 32/P)-ATP binding to the high affinity site on the holoenzyme was examined in the presence of Co/sup + +/, Mn/sup + +/, Mg/sup + +/, and Zn/sup + +/. (..cap alpha../sup 32/P)-ATP-Mg and (..cap alpha../sup 32/P)-ATP-Zn binding to the high-affinity site were compared. Zn-ATP was found to bind to two additional sites on the holoenzyme. The second part of this study investigated the role of cAMP in the dissociation of the native holoenzyme. (..cap alpha../sup 32/P)-ATP-Mg or (..cap alpha../sup 32/P)-ATP-Zn binding to the holoenzyme was followed in the presence of cAMP, and the decrease in binding stoichiometry was taken as indirect evidence of holoenzyme dissociation. Similarly, the role of Mg-ATP in reassociation of the dissimilar subunits was investigated. The slow rate of the biphasic exchange of (/sup 3/H-cAMP) with free cAMP was compared in the presence of various combinations of catalytic subunit, Mg-ATP, Zn-ATP, or the nonhydrolyzable ATP analogues AMPPNP or AMPPCP.
Research Organization:
Virginia Univ., Charlottesville (USA)
OSTI ID:
6985782
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METALS
AMP
ATP
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
CATALYSIS
CATIONS
CHARGED PARTICLES
COBALT
DATA
DAYS LIVING RADIOISOTOPES
ELEMENTS
ENZYMES
EXPERIMENTAL DATA
INFORMATION
IONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
MAGNESIUM
MANGANESE
METALS
NUCLEI
NUCLEOTIDES
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
TRACER TECHNIQUES
TRANSFERASES
TRANSITION ELEMENTS
TRITIUM COMPOUNDS
ZINC