PMID- 10884378
OWN - NLM
STAT- MEDLINE
DCOM- 20001120
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 275
IP  - 42
DP  - 2000 Oct 20
TI  - Phosphorylation by cyclin-dependent protein kinase 5 of the regulatory subunit of 
      retinal cGMP phosphodiesterase. I. Identification of the kinase and its role in 
      the turnoff of phosphodiesterase in vitro.
PG  - 32950-7
AB  - Cyclic GMP phosphodiesterase (PDE) is an essential component in retinal 
      phototransduction. PDE is regulated by Pgamma, the regulatory subunit of PDE, and 
      GTP/Talpha, the GTP-bound alpha subunit of transducin. In previous studies 
      (Tsuboi, S., Matsumoto, H. , Jackson, K. W., Tsujimoto, K., Williamas, T., and 
      Yamazaki, A. (1994) J. Biol. Chem. 269, 15016-15023; Tsuboi, S., Matsumoto, H., 
      and Yamazaki, A. (1994) J. Biol. Chem. 269, 15024-15029), we showed that Pgamma 
      is phosphorylated by a previously unknown kinase (Pgamma kinase) in a 
      GTP-dependent manner in photoreceptor outer segment membranes. We also showed 
      that phosphorylated Pgamma loses its ability to interact with GTP/Talpha, but 
      gains a 10-15 times higher ability to inhibit GTP/Talpha-activated PDE than that 
      of nonphosphorylated Pgamma. Thus, we propose that the Pgamma phosphorylation is 
      probably involved in the recovery phase of phototransduction through shut off of 
      GTP/Talpha-activated PDE. Here we demonstrate that all known Pgammas preserve a 
      consensus motif for cyclin-dependent protein kinase 5 (Cdk5), a protein kinase 
      believed to be involved in neuronal cell development, and that Pgamma kinase is 
      Cdk5 complexed with p35, a neuronal Cdk5 activator. Mutational analysis of Pgamma 
      indicates that all known Pgammas contain a P-X-T-P-R sequence and that this 
      sequence is required for the Pgamma phosphorylation by Pgamma kinase. In three 
      different column chromatographies of a cytosolic fraction of frog photoreceptor 
      outer segments, the Pgamma kinase activity exactly coelutes with Cdk5 and p35. 
      The Pgamma kinase activity ( approximately 85%) is also immunoprecipitated by a 
      Cdk5-specific antibody, and the immunoprecipitate phosphorylates Pgamma. Finally, 
      recombinant Cdk5/p35, which were expressed using clones from a bovine retina cDNA 
      library, phosphorylates Pgamma in frog outer segment membranes in a GTP-dependent 
      manner. These observations suggest that Cdk5 is probably involved in the recovery 
      phase of phototransduction through phosphorylation of Pgamma complexed with 
      GTP/Talpha in mature vertebrate retinal photoreceptors.
FAU - Matsuura, I
AU  - Matsuura I
AD  - Departments of Ophthalmology and Pharmacology, the Kresge Eye Institute, Wayne 
      State University, School of Medicine, Detroit, Michigan 48201, USA.
FAU - Bondarenko, V A
AU  - Bondarenko VA
FAU - Maeda, T
AU  - Maeda T
FAU - Kachi, S
AU  - Kachi S
FAU - Yamazaki, M
AU  - Yamazaki M
FAU - Usukura, J
AU  - Usukura J
FAU - Hayashi, F
AU  - Hayashi F
FAU - Yamazaki, A
AU  - Yamazaki A
LA  - eng
GR  - EY07546/EY/NEI NIH HHS/United States
GR  - EY09631/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Peptide Fragments)
RN  - 0 (Protein Subunits)
RN  - 0 (Recombinant Fusion Proteins)
RN  - EC 2.7.11.1 (Cyclin-Dependent Kinase 5)
RN  - EC 2.7.11.22 (CDK5 protein, human)
RN  - EC 2.7.11.22 (Cdk5 protein, mouse)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinases)
RN  - EC 3.1.4.35 (3',5'-Cyclic-GMP Phosphodiesterases)
RN  - EC 3.6.5.1 (Transducin)
SB  - IM
MH  - 3',5'-Cyclic-GMP Phosphodiesterases/*chemistry/*metabolism
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Cattle
MH  - Cyclin-Dependent Kinase 5
MH  - Cyclin-Dependent Kinases/chemistry/*metabolism
MH  - Humans
MH  - Kinetics
MH  - Mice
MH  - Mutagenesis, Site-Directed
MH  - Peptide Fragments/chemistry
MH  - Phosphorylation
MH  - Protein Subunits
MH  - Ranidae
MH  - Recombinant Fusion Proteins/metabolism
MH  - Retina/*enzymology
MH  - Retinal Cone Photoreceptor Cells/enzymology
MH  - Retinal Rod Photoreceptor Cells/enzymology
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Transducin/metabolism
EDAT- 2000/07/08 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/07/08 11:00
PHST- 2000/07/08 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/07/08 11:00 [entrez]
AID - S0021-9258(20)89186-8 [pii]
AID - 10.1074/jbc.M000702200 [doi]
PST - ppublish
SO  - J Biol Chem. 2000 Oct 20;275(42):32950-7. doi: 10.1074/jbc.M000702200.