title: Phosphosite mapping of HIP-55 protein in mammalian cells.
abstract: In the present study, hematopoietic progenitor kinase 1 (HPK1)-interacting protein of 55 kDa (HIP-55) protein was over-expressed in HEK293 cells, which was genetically attached with 6x His tag. The protein was purified by nickel-charged resin and was then subjected to tryptic digestion. The phosphorylated peptides within the HIP-55 protein were enriched by TiO2 affinity chromatography, followed by mass spectrometry analysis. Fourteen phosphorylation sites along the primary structure of HIP-55 protein were identified, most of which had not been previously reported. Our results indicate that bio-mass spectrometry coupled with manual interpretation can be used to successfully identify the phosphorylation modification in HIP-55 protein in HEK293 cells.
authors.affiliation: Central Laboratory, Jilin University Second Hospital, Changchun 130041, China. liu_ning@jlu.edu.cn.
authors.forename: Ning
authors.initials: N
authors.lastname: Liu
authors.affiliation: Central Laboratory, Jilin University Second Hospital, Changchun 130041, China. sunnn13@mails.jlu.edu.cn.
authors.forename: Ningning
authors.initials: N
authors.lastname: Sun
authors.affiliation: Central Laboratory, Jilin University Second Hospital, Changchun 130041, China. gaoxiang13@mails.jlu.edu.cn.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.affiliation: Institute of Vascular Medicine, Peking University Third Hospital, Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Ministry of Health, Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education and Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing 100191, China. lizijian@bjmu.edu.cn.
authors.forename: Zijian
authors.initials: Z
authors.lastname: Li
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D001665:Binding Sites; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057809:HEK293 Cells; D006639:Histidine / Q000235:genetics; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D010766:Phosphorylation; D018909:src Homology Domains / Q000235:genetics
chemical_list: C117063:DBNL protein, human; D008840:Microfilament Proteins; C033223:polyhistidine; D006639:Histidine