\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
{"_id":"SDzQRpCQBkKH2DatD","bibbaseid":"itoh-fujinoki-kawamura-inaba-ohtake-shimizu-morisawa-textbfpurificationandcharacterizationofthe15kdaproteinfromthespermflagellaofsalmonidtextbffishes-2003","author_short":["Itoh, A.","Fujinoki, M.","Kawamura, T.","Inaba, K.","Ohtake, H.","Shimizu, N.","Morisawa, M."],"bibdata":{"bibtype":"article","type":"article","title":"\\textbfPurification and Characterization of the 15-kDa Protein from the Sperm Flagella of Salmonid \\textbfFishes ","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":[{"propositions":[],"lastnames":["Itoh"],"firstnames":["Atsuko"],"suffixes":[]},{"propositions":[],"lastnames":["Fujinoki"],"firstnames":["Masakatsu"],"suffixes":[]},{"propositions":[],"lastnames":["Kawamura"],"firstnames":["Takeshi"],"suffixes":[]},{"propositions":[],"lastnames":["Inaba"],"firstnames":["Kazuo"],"suffixes":[]},{"propositions":[],"lastnames":["Ohtake"],"firstnames":["Hideki"],"suffixes":[]},{"propositions":[],"lastnames":["Shimizu"],"firstnames":["Nobuyoshi"],"suffixes":[]},{"propositions":[],"lastnames":["Morisawa"],"firstnames":["Masaaki"],"suffixes":[]}],"year":"2003","keywords":"Inaba K","pages":"153–164","bibtex":"@article{itoh_purification_2003,\n\ttitle = {\\textbf{{Purification} and {Characterization} of the 15-{kDa} {Protein} from the {Sperm} {Flagella} of {Salmonid} }\\textbf{{Fishes} }},\n\tvolume = {24},\n\tdoi = {10.2220/biomedres.24.153},\n\tabstract = {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.},\n\tnumber = {4},\n\tjournal = {Biomedical Research},\n\tauthor = {Itoh, Atsuko and Fujinoki, Masakatsu and Kawamura, Takeshi and Inaba, Kazuo and Ohtake, Hideki and Shimizu, Nobuyoshi and Morisawa, Masaaki},\n\tyear = {2003},\n\tkeywords = {Inaba K},\n\tpages = {153--164}\n}\n\n","author_short":["Itoh, A.","Fujinoki, M.","Kawamura, T.","Inaba, K.","Ohtake, H.","Shimizu, N.","Morisawa, M."],"key":"itoh_purification_2003","id":"itoh_purification_2003","bibbaseid":"itoh-fujinoki-kawamura-inaba-ohtake-shimizu-morisawa-textbfpurificationandcharacterizationofthe15kdaproteinfromthespermflagellaofsalmonidtextbffishes-2003","role":"author","urls":{},"keyword":["Inaba K"],"downloads":0,"html":"","metadata":{"authorlinks":{}}},"bibtype":"article","biburl":"http://www.shimoda.tsukuba.ac.jp/smrcbibtext.bib","dataSources":["FmNRw89Cc4gQcNKAj"],"keywords":["inaba k"],"search_terms":["textbfpurification","characterization","kda","protein","sperm","flagella","salmonid","textbffishes","itoh","fujinoki","kawamura","inaba","ohtake","shimizu","morisawa"],"title":"\\textbfPurification and Characterization of the 15-kDa Protein from the Sperm Flagella of Salmonid \\textbfFishes ","year":2003}