Molecular cloning and expression of a mouse brain cDNA encoding a novel protein target of calcyclin.
A protein target of mouse calcyclin, p30, which we call calcyclin-binding protein (CacyBP), was identified in mouse brain and Ehrlich ascites tumor (EAT) cells. The amino acid sequence of the CacyBP chymotryptic peptide was used to prepare synthetic oligonucleotides that served as a probe to screen the mouse brain cDNA library. A 1.4-kb positive clone was detected, isolated, and sequenced. The analyzed clone contains an open reading frame encoding a protein of a molecular mass of approximately 26 kDa. The nucleotide and predicted amino acid sequences indicate that CacyBP is a novel protein. The results obtained from northern blots show that the CacyBP gene is expressed predominantly in mouse brain and EAT cells. Using a pGEX vector the recombinant CacyBP was expressed in Escherichia coli, and its properties were analyzed. The recombinant protein interacts with calcyclin at a physiologically relevant range of Ca2+ in solution during affinity chromatography and on blots. Because CacyBP, like calcyclin, is present in the brain, the interaction of these two proteins might be involved in calcium signaling pathways in neuronal tissue.
Filipek
A
A
Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology, Warsaw, Poland.
Kuźnicki
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism / Q000473:pathology; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; D014407:Tumor Cells, Cultured / Q000378:metabolism
C491778:CACYBP protein, human; C111922:Cacybp protein, mouse; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D011994:Recombinant Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C489134:S100a6 protein, mouse; D002118:Calcium