Phosphosite Mapping of HIP-55 Protein in Mammalian Cells

International Journal of Molecular Sciences

Ning Liu¹  Ningning Sun¹  Xiang Gao¹ 

[1] Central Laboratory, Jilin University Second Hospital, Changchun 130041, China; E-Mails:

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

DOI  :  10.3390/ijms15034903

来源: mdpi

【 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】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland