Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry

doi:10.1016/S1671-2927(00)8562

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (3): 439-445.DOI: 10.1016/S1671-2927(00)8562

Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry

LI Shan-shan, WEI Hong-yang, BU Deng-pan, ZHANG Le-ying, ZHOU Ling-yun

1.State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China

收稿日期:2010-12-10 出版日期:2012-03-01 发布日期:2012-03-11
通讯作者: Correspondence WANG Jia-qi, Tel: +86-10-62890458, Fax: +86-10-62897587, E-mail: jqwangcaas@gmail.com
作者简介:LI Shan-shan, E-mail: leeshanshanshan@163.com;
基金资助:
The project was supported by an earmark fund for the National Key Basic Research Program of China (2011CB100805).

Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry

LI Shan-shan, WEI Hong-yang, BU Deng-pan, ZHANG Le-ying, ZHOU Ling-yun

1.State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China

Received:2010-12-10 Online:2012-03-01 Published:2012-03-11
Contact: Correspondence WANG Jia-qi, Tel: +86-10-62890458, Fax: +86-10-62897587, E-mail: jqwangcaas@gmail.com
About author:LI Shan-shan, E-mail: leeshanshanshan@163.com;
Supported by:
The project was supported by an earmark fund for the National Key Basic Research Program of China (2011CB100805).

摘要/Abstract

摘要： Protein phosphorylation is an important post-translational modification that regulates milk protein structure and function. The objective of this study was to analyze the presence of phosphorylated casein. Bovine milk proteins were first separated by SDS polyacrylamide gel electrophoresis. After in gels digestion and extraction, phosphorylated peptides were enriched by titanium dioxide and identified by ultra performance liquid chromatography coupled with nano electrospray ionization tandem mass spectrometry. This method ensured the identification of 20 phosphorylated peptides, including 7 phosphorylated forms of αs1-casein, 8 αs2-casein, and 5 β-casein. Eight phosphorylated sites derived from 3 αs1-caseins, 3 αs2-caseins, and 2 β-caseins were also identified, and localized on residues Ser61, Ser63 and Ser130 in αs1-casein; Thr145, Ser146 and Ser158 in αs2-casein; and Ser50 and Thr56 in β-casein. These findings provide valuable information for investigating casein phosphorylation of the bovine milk.

关键词: bovine milk casein, phosphorylation, mass spectrometry, titanium dioxide enrichment

Abstract: Protein phosphorylation is an important post-translational modification that regulates milk protein structure and function. The objective of this study was to analyze the presence of phosphorylated casein. Bovine milk proteins were first separated by SDS polyacrylamide gel electrophoresis. After in gels digestion and extraction, phosphorylated peptides were enriched by titanium dioxide and identified by ultra performance liquid chromatography coupled with nano electrospray ionization tandem mass spectrometry. This method ensured the identification of 20 phosphorylated peptides, including 7 phosphorylated forms of αs1-casein, 8 αs2-casein, and 5 β-casein. Eight phosphorylated sites derived from 3 αs1-caseins, 3 αs2-caseins, and 2 β-caseins were also identified, and localized on residues Ser61, Ser63 and Ser130 in αs1-casein; Thr145, Ser146 and Ser158 in αs2-casein; and Ser50 and Thr56 in β-casein. These findings provide valuable information for investigating casein phosphorylation of the bovine milk.

Key words: bovine milk casein, phosphorylation, mass spectrometry, titanium dioxide enrichment

LI Shan-shan, WEI Hong-yang, BU Deng-pan, ZHANG Le-ying, ZHOU Ling-yun . Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry[J]. Journal of Integrative Agriculture, 2012, 12(3): 439-445.

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Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry

Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry

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