PMID- 7559450
OWN - NLM
STAT- MEDLINE
DCOM- 19951106
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 270
IP  - 39
DP  - 1995 Sep 29
TI  - Characterization of active and inactive forms of the JAK2 protein-tyrosine kinase 
      produced via the baculovirus expression vector system.
PG  - 23084-9
AB  - Three forms of rat JAK2 (type 2 Janus tyrosine kinase) were produced via the 
      baculovirus expression vector system. Recombinant baculoviruses encoded either 
      the full-length rat jak2 cloned from the Nb2-SP cell line (rJAK2), a 
      carboxyl-terminal deletion mutant lacking the putative catalytic domain (rJAK2(C 
      delta 795)), or an amino-terminal deletion mutant containing the putative 
      catalytic domain ((N delta 661)rJAK2). The proteins produced in infected Sf21 
      cells were assayed for phosphotyrosine content and autophosphorylating activity. 
      Tyrosine phosphorylation of rJAK2 was not observed 1 day postinfection when rJAK2 
      was initially produced but was apparent 2 or more days postinfection when the 
      rJAK2 level had significantly increased. Tyrosine phosphorylation of rJAK2(C 
      delta 795) was not observed; further, coproduction of rJAK2(C delta 795) with 
      rJAK2 blocked tyrosine phosphorylation of rJAK2, consistent with previously 
      published results (Zhuang, H., Patel, S. V., He, T-C., Sonsteby, S. K., Niu, Z., 
      and Wojchowski, D. M. (1994) J. Biol. Chem. 269, 21411-21414). Mutant (N delta 
      661)rJAK2 exhibited a robust tyrosine phosphorylation signal. A second 62-kDa 
      tyrosine phosphoprotein co-immunoprecipitated with (N delta 661)rJAK2 but not 
      with rJAK2 or rJAK2(C delta 795). Both rJAK2 and (N delta 661)rJAK2 incorporated 
      phosphate under in vitro kinase assay conditions, but rJAK2(C delta 795) did not. 
      A JAK2 oligomer with interacting catalytic sites and/or inhibitory sites would 
      provide a simple model to describe these results.
FAU - Duhe, R J
AU  - Duhe RJ
AD  - Biological Carcinogenesis and Development Program, Science Applications 
      International Corporation, Frederick, Maryland 21702-1201, USA.
FAU - Farrar, W L
AU  - Farrar WL
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 21820-51-9 (Phosphotyrosine)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.2 (Jak2 protein, rat)
RN  - EC 2.7.10.2 (Janus Kinase 2)
SB  - IM
MH  - Animals
MH  - Baculoviridae
MH  - Blotting, Western
MH  - Cloning, Molecular
MH  - Janus Kinase 2
MH  - Kinetics
MH  - Mutagenesis
MH  - Phosphotyrosine/analysis
MH  - Protein-Tyrosine Kinases/analysis/biosynthesis/*metabolism
MH  - *Proto-Oncogene Proteins
MH  - Rats
MH  - Recombinant Proteins/analysis/biosynthesis/metabolism
MH  - Sequence Deletion
MH  - Signal Transduction
MH  - Spodoptera
MH  - Time Factors
MH  - Transfection
EDAT- 1995/09/29 00:00
MHDA- 1995/09/29 00:01
CRDT- 1995/09/29 00:00
PHST- 1995/09/29 00:00 [pubmed]
PHST- 1995/09/29 00:01 [medline]
PHST- 1995/09/29 00:00 [entrez]
AID - S0021-9258(18)90072-4 [pii]
AID - 10.1074/jbc.270.39.23084 [doi]
PST - ppublish
SO  - J Biol Chem. 1995 Sep 29;270(39):23084-9. doi: 10.1074/jbc.270.39.23084.