Site-directed mutation makes rabbit calcyclin dimer.
Unlike human, rat and mouse calcyclin, purified rabbit calcyclin did not form a dimer on Tricine SDS-PAGE under non-reduced conditions. Based on the internal peptide sequence of rabbit calcylin, we isolated and sequenced a cDNA clone encoding calcyclin. The sequence of this clone (pCalC) is 629 bp long and codes 90 amino acid residues of a protein with a molecular mass of 10,153 Da. By Northern blot analysis, a major band of 0.9 kbp and a minor band of 2.6 kbp were detected in the lung. The recombinant calcyclin mutated serine at the third position to cysteine was expressed in E. coli and made dimer formation under non-reduced conditions on SDS-PAGE. Whether or not this type of mutation which prevents dimer formation of calcyclin plays a physiological role in the rabbit lung is the subject of an ongoing study.
Ando
Y
Y
Department of Pharmacology, Nagoya University School of Medicine, Japan.
Watanabe
M
M
Akatsuka
H
H
Tokumitsu
H
H
Hidaka
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular; D003545:Cysteine; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D008168:Lung; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; D017386:Sequence Homology, Amino Acid
D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D011994:Recombinant Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C489134:S100a6 protein, mouse; C489132:S100a6 protein, rat; D003545:Cysteine