Use of Titanium Dioxide To Find Phosphopeptide and Total Protein Changes During Epididymal Sperm Maturation

作者：Baker Mark A^*; Smith Nathan D; Hetherington Louise; Pelzing Matthias; Condina Mark R; Aitken R John

来源：Journal of Proteome Research, 2011, 10(3): 1004-1017.

DOI：10.1021/pr1007224

• sperm phosphorylation
• epididymal maturation
• mass spectrometry
• proteomics
• titanium dioxide
• label-free quantitation
• ornathine decarboxylase
• heat shock protein 90

Although the overall performance of modern mass spectrometers has increased, proteomic analysis of complex samples still requires prefractionation either at the protein or peptide level to allow for in-depth analysis of normal cellular function. Here, we report a novel way to identify protein changes occurring during sperm development through the epididymis. Phosphopeptides were first enriched from either the rat caput or caudal regions of the epididymides using TiO(2), and the profiles then quantitatively compared. We show that 77 TiO(2)-enriched peptides become Through the use of immunoblot analysis, we confirmed that three significantly modified in the epididymis, equating to 53 proteins. proteins, ornithine-decarboxylase antizyme 3, heat-shock protein 90 alpha, and testis-lipid binding protein, undergo major protein loss during epididymal passage. Many other proteins, including t-complex protein 10 and Spatal8 show testis unique expression, appear to undergo phosphorylation during this same time frame. These data provide mechanistic insight into the means by which spermatozoa acquire functionality during epididymal transit.

• 出版日期2011-3