Expression in Escherichia coli and phosphorylation with cAMP-dependent protein kinase of the N-terminal region of human endothelial actin-binding protein.
The N-terminal region of human endothelial actin-binding protein was subcloned and expressed in the pT 7-7/E. coli BL 21 (DE 3) system. This peptide was efficiently expressed in E. coli as indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by western analysis using a monoclonal antibody raised against this part of the molecule. As predicted by the amino acid sequence this truncated protein (residues 21-1569), corresponding to 164 KD and containing the actin-binding domain near the amino-terminus, could be phosphorylated by cAMP-dependent protein kinase unmasking a phosphorylation site which is not apparent in the native ABP protein.
Jay
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Department of Biochemistry, State University of New York, Health Science Center at Brooklyn 11203.
Stracher
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D000199:Actins / Q000378:metabolism; D001665:Binding Sites; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D018076:DNA, Complementary; D004727:Endothelium / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D013912:Threonine / Q000378:metabolism
D000199:Actins; D018076:DNA, Complementary; D008840:Microfilament Proteins; D010446:Peptide Fragments; D013912:Threonine; D017868:Cyclic AMP-Dependent Protein Kinases