PMID- 8550544
OWN - NLM
STAT- MEDLINE
DCOM- 19960220
LR  - 20211203
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 271
IP  - 1
DP  - 1996 Jan 5
TI  - Induced release of cell surface protein kinase yields CK1- and CK2-like enzymes 
      in tandem.
PG  - 111-9
AB  - Several types of cell exhibit cell surface protein kinase (ecto-PK) activities 
      with Ser/Thr-specificity. Ecto-PK sharing certain characteristics of protein 
      kinase CK2 can be detached from intact cells by interaction with exogenous 
      substrates (Kubler, D., Pyerin, W., Burow, E., and Kinzel, V. (1983) Proc. Natl. 
      Acad. Sci. U.S.A. 80, 4021-4025). However, a detailed molecular analysis of this 
      ecto-PK was hampered by the vanishingly small amounts of labile enzyme protein 
      obtained by substrate-inducible enzyme release. We now describe the stabilization 
      and enrichment of released ecto-PK by precipitation with polyethylene glycol 
      followed by affinity chromatography on heparin-agarose. Ecto-PK is shown to 
      consist of two separate forms released in tandem, ecto-PK I and ecto-PK II. 
      Comparison with cell homogenates as well as cell surface biotinylation 
      experiments excluded contamination with intracellular PK. Purified ecto-PK I and 
      ecto-PK II exhibit respectively selective phosphorylation of CK1- and 
      CK2-specific peptide substrates, a complementary sensitivity to inhibitory agents 
      and a differential use of the cosubstrates ATP and GTP. Ecto-PK I consists of a 
      40-kDa moiety; the ecto-PK II is an ensemble of three components of 43- and 
      40-kDa (catalytic subunits) and a noncatalytic 28-kDa subunit. In addition, 
      components of the ecto-PK II react with CK2-specific antibodies. Further, 
      comparative peptide mapping and the results of mass spectrometry in combination 
      with assignment of amino acid sequences confirmed that ecto-PK II is closely 
      related if not identical to the protein kinase CK2. Assays with intact cells that 
      result in the phosphorylation of a variety of endogenous membrane proteins showed 
      that both ecto-PKs participate, and further, certain ecto-PK substrates become 
      preferentially labeled by one or another of the enzymes, whereas others are 
      phosphorylated by both ecto-PK activities.
FAU - Walter, J
AU  - Walter J
AD  - Department of Pathochemistry, German Cancer Research Center (Deutsches 
      Krebsforschungszentrum, Heidelberg, Federal Republic of Germany.
FAU - Schnolzer, M
AU  - Schnolzer M
FAU - Pyerin, W
AU  - Pyerin W
FAU - Kinzel, V
AU  - Kinzel V
FAU - Kubler, D
AU  - Kubler D
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 6SO6U10H04 (Biotin)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.11.1 (Casein Kinase II)
RN  - EC 2.7.11.1 (Casein Kinases)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Biotin/metabolism
MH  - Blotting, Western
MH  - Casein Kinase II
MH  - Casein Kinases
MH  - Cell Membrane/enzymology
MH  - Chromatography, Affinity
MH  - HeLa Cells
MH  - Humans
MH  - Molecular Sequence Data
MH  - Phosphorylation
MH  - Protein Kinases/isolation & purification/*metabolism
MH  - Protein Serine-Threonine Kinases/isolation & purification/*metabolism
MH  - Substrate Specificity
EDAT- 1996/01/05 00:00
MHDA- 1996/01/05 00:01
CRDT- 1996/01/05 00:00
PHST- 1996/01/05 00:00 [pubmed]
PHST- 1996/01/05 00:01 [medline]
PHST- 1996/01/05 00:00 [entrez]
AID - S0021-9258(18)95311-1 [pii]
AID - 10.1074/jbc.271.1.111 [doi]
PST - ppublish
SO  - J Biol Chem. 1996 Jan 5;271(1):111-9. doi: 10.1074/jbc.271.1.111.