p30, a novel protein target of mouse calcyclin (S100A6).
A novel protein target of mouse calcyclin (S100A6) was detected by a gel overlay method with 125I-labelled calcyclin. Interaction of calcyclin with its 30 kDa target protein (p30) present in Ehrlich ascites tumour (EAT) cells depended on the presence of Ca2+ ions. The binding of p30, evidenced by the reaction with 125I-labelled calcyclin, was found to be of higher affinity than the binding between mouse calcyclin and annexin II or glyceraldehyde-3-phosphate dehydrogenase. Examination of tissue extracts by the gel overlay method has shown that p30 is present not only in the EAT cells but also in mouse brain and spleen. This novel target protein of mouse calcyclin was purified to homogeneity from EAT cells by means of Phenyl-Sepharose chromatography, affinity chromatography and CM-cellulose chromatography. Purified p30 was digested with alpha-chymotrypsin and a partial amino acid sequence of one of the resulting peptides was established. A database search analysis revealed that the sequence is unique, with a similarity of less than 55% to any other known protein sequence.
Filipek
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A
Department of Muscle Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland.
Wojda
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U
D000595:Amino Acid Sequence; D000818:Animals; D017306:Annexin A2 / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism; D018797:Cell Cycle Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; D014407:Tumor Cells, Cultured
D017306:Annexin A2; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C489134:S100a6 protein, mouse; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases