Int. J. Mol. Sci. 2014, 15(3), 4903-4914; https://doi.org/10.3390/ijms15034903
Phosphosite Mapping of HIP-55 Protein in Mammalian Cells
1
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
2
Central Laboratory, Jilin University Second Hospital, Changchun 130041, China
*
Author to whom correspondence should be addressed.
Received: 19 January 2014 / Revised: 20 February 2014 / Accepted: 7 March 2014 / Published: 19 March 2014
(This article belongs to the Special Issue Mass Spectrometry Application in Biology)
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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. View Full-TextKeywords:
mass spectrometry; HIP-55; phosphorylation
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Liu, N.; Sun, N.; Gao, X.; Li, Z. Phosphosite Mapping of HIP-55 Protein in Mammalian Cells. Int. J. Mol. Sci. 2014, 15, 4903-4914.
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