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Mol. Hum. Reprod. Advance Access published online on April 3, 2009
Molecular Human Reproduction, doi:10.1093/molehr/gap028
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Mass spectrometry analysis of dynamic post-translational modifications of TH2B during spermatogenesis

Shuang Lu1,2, Yong M. Xie4,*, Xin Li2, Ji Luo5, Xin Q. Shi3, Xin Hong4, Ying H. Pan5 and Xu Ma1,2,*

1Graduate School, Peking Union Medical College, Beijing, China 2Genetics Department of the National Institute Research for Family Planning; WHO Human Reproduction Research Collaborative Center, Beijing, China 3Endocrine Department of the National Institute Research for Family Planning 4Bejing Office, Bruker Daltonics Inc., Beijing, China 5 Institute of Crop Science, Chinese Academy of Agricultural Sciences/The National Key Facility for Crop Gene Resources and Genetic Improvement, NFCRI, Beijing, China

* To whom correspondence should be addressed. X.Ma, E-mail:genetic{at}263.net.cn and Y. Xie, yongming.xie{at}bruker.com.cn

TH2B, an important testis histone, plays a key role in remodeling chromatin structure during spermatogenesis. We present a detailed study of post-translational modifications (PTMs) of histone TH2B from different developmental stages of sperm cells, using a combination of high performance liquid chromatography (HPLC), enzymatic Glu-c digestions of peptides, liquid chromatography – mass spectrometry (LC-MS) and LC-MS/MS analysis. The results showed modification patterns of the intact histone TH2B during spermatogenesis. Acetylated TH2B was most abundant in spermatogonia (28.9%) as compared with the spermatocytes (8.3%) and round spermatids (11.2%). Several new PTMs of TH2B were identified. In spermatogonia, spermatocytes, and round spermatids, T116 and K117 were modified by phosphorylation and methylation, respectively, forming a novel "phospho switch" site. The identified modification patterns of histone TH2B in spermatogenic cells provides a basis for future studies on histone coding and epigenetic regulation during spermatogenesis.

Key Words: spermatogenesis/post-translational modifications/chromatin/testis-specific histones/TH2B

Submitted on February 11, 2009; resubmitted on March 11, 2009; accepted on March 31, 2009.


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