PMID- 6573204
OWN - NLM
STAT- MEDLINE
DCOM- 19830617
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 22
IP  - 6
DP  - 1983 Mar 15
TI  - Identification of the catalytic subunit of an oligomeric casein kinase (G type). 
      Affinity labeling of the nucleotide site using 
      5'-[p-(fluorosulfonyl)benzoyl]adenosine.
PG  - 1452-9
AB  - Identification of the catalytic subunit of a G type [using guanosine 
      5'-triphosphate (GTP) as well as adenosine 5'-triphosphate (ATP) as phosphate 
      donor], oligomeric, cyclic nucleotide independent casein kinase purified from 
      bovine lung was carried out after reaction with 
      5'-[p-(fluorosulfonyl)-benzoyl]adenosine (FSBA) and isolation of the subunit 
      components of the enzyme. FSBA exhibited the major characteristics of an affinity 
      label reacting at the nucleotide (ATP, GTP) site of the casein kinase. FSBA acted 
      as a competitive inhibitor of ATP (and GTP), led to complete inactivation of the 
      enzyme in a reaction showing two kinetic steps, and became irreversibly bound to 
      the protein. After being labeled with FSBA, the casein kinase (apparent molecular 
      weight of 140 000) was separated into its two monomeric components of apparent 
      molecular weights 38 000 (alpha) and 27 000 (beta), respectively, after sodium 
      dodecyl sulfate-polyacrylamide gel electrophoresis. Use of radioactive FSBA 
      showed that specific affinity labeling was limited to the alpha casein kinase 
      subunit. This result was in agreement with the fact that casein kinase activity 
      was found associated with the alpha monomer after electrophoretic separation of 
      the alpha and beta subunits. It may thus be concluded that the largest (alpha) 
      subunit contains the catalytic site of the casein kinase G. Electrophoretic 
      analysis of purified protein kinase under denaturing conditions suggested an 
      alpha 3 beta 2 combination for an apparent molecular weight of 130 000-140 000. 
      However, a maximum of 2 mol of FSBA could be specifically bound to the alpha 
      subunit per mol of enzyme, with a concomitant complete inactivation. These data 
      would be in agreement with an alpha 2 beta 2 subunit composition for casein 
      kinase G, as proposed by other research groups for a similar type of protein 
      kinase of different sources. These observations suggest that the alpha subunits 
      are functionally similar, each of them containing a nucleotide (ATP, GTP) binding 
      site. The possible role of the beta subunit in the enzyme activity remains to be 
      established.
FAU - Feige, J J
AU  - Feige JJ
FAU - Cochet, C
AU  - Cochet C
FAU - Pirollet, F
AU  - Pirollet F
FAU - Chambaz, E M
AU  - Chambaz EM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Affinity Labels)
RN  - 0 (Nucleotides)
RN  - 0 (Protein Kinase Inhibitors)
RN  - 78859-42-4 (5'-(4-fluorosulfonylbenzoyl)adenosine)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.11.1 (Casein Kinases)
RN  - K72T3FS567 (Adenosine)
SB  - IM
MH  - Adenosine/*analogs & derivatives
MH  - Affinity Labels
MH  - Animals
MH  - Binding Sites
MH  - Casein Kinases
MH  - Catalysis
MH  - Cattle
MH  - Chemical Phenomena
MH  - Chemistry
MH  - In Vitro Techniques
MH  - Kinetics
MH  - Nucleotides/metabolism
MH  - Protein Kinase Inhibitors
MH  - *Protein Kinases/metabolism
EDAT- 1983/03/15 00:00
MHDA- 1983/03/15 00:01
CRDT- 1983/03/15 00:00
PHST- 1983/03/15 00:00 [pubmed]
PHST- 1983/03/15 00:01 [medline]
PHST- 1983/03/15 00:00 [entrez]
AID - 10.1021/bi00275a020 [doi]
PST - ppublish
SO  - Biochemistry. 1983 Mar 15;22(6):1452-9. doi: 10.1021/bi00275a020.