Site-Specific Proteomics Approach for Study Protein S-Nitrosylation

Journal Article · Wed Sep 01 00:00:00 EDT 2010 · Analytical Chemistry, 82(17):7160-7168

DOI:https://doi.org/10.1021/ac100569d· OSTI ID:990126

Liu, Miao; Hou, Jinxuan; Huang, Lin; Huang, Xin; Heibeck, Tyler H; Zhao, Rui; Pasa-Tolic, Ljiljana; Smith, Richard D; Li, Yan; Fu, Kai; Zhang, Zhixin; Hinrichs, Steven; Ding, Shi-Jian

Here we present a novel and robust method for the identification of protein S-nitrosylation sites in complex protein mixtures. The approach utilizes the cysteinyl affinity resin to selectively enrich S-nitrosylated peptides reduced by ascorbate followed by nanoscale liquid chromatography tandem mass spectrometry. Two alkylation agents with different added masses were employed to differentiate the S-nitrosylation sites from the non-Snitrosylation sites. We applied this approach to MDA-MB-231 cells treated with Angeli’s salt, a nitric oxide donor that has been shown to inhibit breast tumor growth and angiogenesis. A total of 162 S-nitrosylation sites were identified and an S-nitrosylation motif was revealed in our study. The 162 sites are significantly more than the number reported by previous methods, demonstrating the efficiency of our approach. Our approach will further facilitate the functional study of protein S-nitrosylation in cellular processes and may reveal new therapeutic targets.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 990126
• Report Number(s): PNNL-SA-75223; 32893; TRN: US201020%%136
• Journal Information: Analytical Chemistry, 82(17):7160-7168, Vol. 82, Issue 17
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
AFFINITY
ALKYLATION
CHROMATOGRAPHY
EFFICIENCY
FUNCTIONALS
MAMMARY GLANDS
MASS SPECTROSCOPY
MIXTURES
NEOPLASMS
NITRIC OXIDE
PEPTIDES
PROTEINS
RESINS
TARGETS
Environmental Molecular Sciences Laboratory