Beyond linker histones and high mobility group proteins: global profiling of perchloric acid soluble proteins.
Extraction with HClO(4) provides an easy method for efficient enrichment of both histone H1 and HMG proteins from a variety of tissues. Usually, the histone and the HMG proteins are the most abundant components of the extracts, however, other proteins have frequently been observed but only seldom studied in more detail. Here we describe a study aimed at global characterization of HClO(4) extractable proteins from breast cancer cell lines. We report identification of 150 unique proteins by liquid chromatography tandem mass spectrometry including almost all major histone H1 variants and canonical members of the HMG protein families. In the extracts, diverse proteins with HMG-like amino acid composition were identified and their post-translational modifications were mapped. Importantly, those include multiple proteins known or supposed to be related to cell proliferation and cancer. Since purification of these proteins as well as low abundant variants of histone and HMG proteins is difficult due to their metabolic instability, characterization of these proteins from crude extracts can facilitate studies aimed at better understanding of their function.
Zougman
Alexandre
A
Protana Inc., Toronto, Ontario, Canada M9W 7H4.
Wiśniewski
Jacek R
JR
D000595:Amino Acid Sequence; D001943:Breast Neoplasms / Q000473:pathology; D002457:Cell Extracts / Q000737:chemistry; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D014644:Genetic Variation; D006609:High Mobility Group Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006657:Histones / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010472:Perchlorates / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D012995:Solubility
D002457:Cell Extracts; D006609:High Mobility Group Proteins; D006657:Histones; D010472:Perchlorates; D020543:Proteome