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Please use this identifier to cite or link to this item: http://dspace.library.iitb.ac.in/jspui/handle/10054/10255

Title: Purification and characterization of a type II phosphatidylinositol 4-kinase from rat spleen and comparison with a PtdIns 4-kinase from lymphocytes
Authors: VERGHESE, M
FERNANDIS, AZ
SUBRAHMANYAM, G
Keywords: t-cell activation
antigen receptor
signal-transduction
a431 cells
phosphoinositide kinases
tyrosine phosphorylation
protein-kinase
membranes
stimulation
turnover
Issue Date: 1999
Publisher: NATL INST SCIENCE COMMUNICATION
Citation: INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS, 36(1), 1-9
Abstract: A PtdIns 4-kinase from rat spleen particulate fraction was purified to homogeneity and its molecular properties were compared with a PtdIns 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 PtdIns 4-kinase. Polyclonal antibodies raised against PtdIns 4-kinase inhibited the enzyme activity in in vitro assays. Analysis of adult rat tissue particulate fractions on immunoblots showed restricted immunoreactivity among PtdIns 4-kinases. However, the immunoreactivity is conserved in lymphoid tissues from mouse to human, suggesting that lymphoid tissue has a distinct PtdIns 4-kinase. Activation of rat splenocytes with Con A showed two fold increase in PtdIns 4-kinase activity. Comparison of PtdIns 4-kinases from spleen and splenic lymphocytes showed identical chromatographic behaviour, molecular mass, immunoreactivity, K-m values for PtdIns and inhibition by adenosine.
URI: http://dspace.library.iitb.ac.in/xmlui/handle/10054/10255
http://hdl.handle.net/10054/10255
ISSN: 0301-1208
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