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Purification and characterization of a type II phosphatidylinositol 4-kinase from rat spleen and comparison with a Ptdlns 4-kinase from lymphocytes

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Title Purification and characterization of a type II phosphatidylinositol 4-kinase from rat spleen and comparison with a Ptdlns 4-kinase from lymphocytes
 
Creator Verghese, Mary
Fernandis, Aaron Z
Subrahmanyam, Gosukonda
 
Description 1-9
A Ptdlns 4-kinase from rat spleen particulate fraction was purified
to homogeneity and its molecular properties were compared with a Ptdlns
4-kinase from splenic lymphocytes. The enzyme activity was solubilized from
spleen particulate

fraction with Triton X-100 and chromatographed sequentially on
phosphocellulose, DEAE-sephacel, heparin acrylamide and hydroxyapatite columns.
The purified enzyme preparation showed a 55 kDa band on SDS-PAGE with silver
staining. Renaturation of the enzyme activity from SDS-PAGE showed that it
comigrated with the 55 kDa protein. Characterization of the enzyme showed that
it was a type II Ptdlns 4-kinase. Polyclonal antibodies raised against Ptdlns
4-kinase inhibited the enzyme activity in in vitro assays. Analysis of
adult rat tissue particulate fractions on immunoblots showed restricted immunoreactivity
among Ptdlns 4-kinases. However, the immunoreactivity is conserved in lymphoid
tIssues from mouse to human, suggesting that lymphoid tissue has a distinct
Ptdlns 4-kinase. Activation of rat splenocytes with Con A showed two fold in
crease in Ptdlns 4-kinase activity. Comparison of Ptdlns 4-kinases from spleen
and splenic lymphocytes showed

identical chromatographic behaviour, molecular
mass, immunoreactivity, Km values for Ptdlns and inhibition
by adenosine.
 
Date 2012-12-30T20:23:10Z
2012-12-30T20:23:10Z
1999-02
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/15408
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.36(1) [February 1999]