Expression in Escherichia coli, phosphorylation with cAMP-dependent protein kinase and proteolysis by calpain of a 71-kDa domain of human endothelial actin binding protein.
A middle region of human endothelial actin-binding protein (ABP) was subcloned and expressed in the pT7-7/E. coli BL21 (DE3) system. As predicted by the amino acid sequence this 71 kD truncated protein (residues 1717-2360) contained a calpain cleavage site and two of the three presumptive cAMP-dependent protein kinase phosphorylation sites. This peptide fragment comprised all the elements needed to confer stability against calpain proteolysis to ABP after PKA phosphorylation.
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Departamento de Cultivo de Tejidos, Instituto Nacional de Cardiologia, Mexico, Distrito Federal, Mexico.
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D001665:Binding Sites; D002154:Calpain / Q000502:physiology*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D006868:Hydrolysis; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D010766:Phosphorylation; D017434:Protein Structure, Tertiary
D002352:Carrier Proteins; C034037:F-actin-binding proteins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D017868:Cyclic AMP-Dependent Protein Kinases; D002154:Calpain