\textbfPurification and Characterization of the 15-kDa Protein from the Sperm Flagella of Salmonid \textbfFishes . Itoh, A., Fujinoki, M., Kawamura, T., Inaba, K., Ohtake, H., Shimizu, N., & Morisawa, M. Biomedical Research, 24(4):153–164, 2003.
doi  abstract   bibtex   
Cyclic AMP-dependent phosphorylation of tyrosine residues in a protein with a molecular mass of 15-kDa is thought to play a key role in the initiation of sperm motility in salmonid fishes (7). In this study, flagellar proteins were solubilized with 7 M urea and fractionated by sucrose densitygradient isoelectrie focusing. A band relevant to 15-kDa was detected in the fraction at pH 5 on electrophoresis and analyzed using phosphotyrosine antibody. Analysis of the band by reversed phase chromatography showed a single peak, suggesting that the 15-kDa protein was completely purified. Amino acid sequence of His—Ile-Phe at the N-terminus was detected by analysis using peptide sequencer, and the fourth base could not be determined. Further analysis of the molecular structure of the protein by LC—MS/MS showed that partial amino acid sequence of the 15-kDa protein from tryptic fragments were similar to parts of the amino acid sequence of tubulin. The protein was not detected in inununoblots using anti-tubulin polyclonal antibody, suggesting that the 15-kDa protein may be not debris of the tubulin but a novel protein with a common partial sequence of tubulin.
@article{itoh_purification_2003,
	title = {\textbf{{Purification} and {Characterization} of the 15-{kDa} {Protein} from the {Sperm} {Flagella} of {Salmonid} }\textbf{{Fishes} }},
	volume = {24},
	doi = {10.2220/biomedres.24.153},
	abstract = {Cyclic AMP-dependent phosphorylation of tyrosine residues in a protein with a molecular mass of 15-kDa is thought to play a key role in the initiation of sperm motility in salmonid fishes (7). In this study, flagellar proteins were solubilized with 7 M urea and fractionated by sucrose densitygradient isoelectrie focusing. A band relevant to 15-kDa was detected in the fraction at pH 5 on electrophoresis and analyzed using phosphotyrosine antibody. Analysis of the band by reversed phase chromatography showed a single peak, suggesting that the 15-kDa protein was completely purified. Amino acid sequence of His—Ile-Phe at the N-terminus was detected by analysis using peptide sequencer, and the fourth base could not be determined. Further analysis of the molecular structure of the protein by LC—MS/MS showed that partial amino acid sequence of the 15-kDa protein from tryptic fragments were similar to parts of the amino acid sequence of tubulin. The protein was not detected in inununoblots using anti-tubulin polyclonal antibody, suggesting that the 15-kDa protein may be not debris of the tubulin but a novel protein with a common partial sequence of tubulin.},
	number = {4},
	journal = {Biomedical Research},
	author = {Itoh, Atsuko and Fujinoki, Masakatsu and Kawamura, Takeshi and Inaba, Kazuo and Ohtake, Hideki and Shimizu, Nobuyoshi and Morisawa, Masaaki},
	year = {2003},
	keywords = {Inaba K},
	pages = {153--164}
}

Downloads: 0