Phosphosite Mapping of HIP-55 Protein in Mammalian Cells

International Journal of Molecular Sciences

Ning Liu¹  Xiang Gao¹  Ningning Sun¹  Zijian Li² 

[1] Central Laboratory, Jilin University Second Hospital, Changchun 130041, China;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;

关键词: mass spectrometry;    HIP-55;    phosphorylation;   

DOI  :  10.3390/ijms15034903

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown