A reference map and identification of porcine testis proteins using 2-DE and MS

Abstract

The development of the testis is essential for maturation of male mammals. A complete understanding of proteins expressed in the testis will provide biological information on many reproductive dysfunctions in males. The purposes of this study were to apply a proteomic approach to investigating protein composition and to establish a 2-D PAGE reference map for porcine testis proteins. MALDI-TOF MS was performed for protein identification. When 1 mg of total proteins was assayed by 2-D PAGE and stained with colloidal CBB, more than 400 proteins with a pI of pH 3-10 and M(r) of 10-200 kDa could be detected. Protein expression varied among individuals, with CV between 4.7 and 131.5%. A total of 447 protein spots were excised for identification, among which 337 spots were identified by searching the mass spectra against the NCBInr database. Identification of the remaining 110 spots was unsuccessful. A 2-D PAGE-based porcine testis protein database has been constructed on the basis of the results and will be published on the WWW. This database should be valuable for investigating the developmental biology and pathology of porcine testis.

A proteome reference map and virulence factors analysis of Yersinia pestis 91001

“…Reference map of proteome has been established for many bacteria, including some important pathogens [13][14][15][16][17][18][19][20][21][22][23][24][25][26]. The 2-DE proteome database focused on pathogenic bacteria has been established since 1999 [27].…”

An integrated bioinformatics analysis of mouse testis protein profiles with new understanding

“…The proteomics of mammal testis should be the first effort to describe these processes and deal with testis-specific proteins to explore their structures and functions as well as their implicated clinical applications. However, fewer related works were done systematically, most of which focused mainly on the description of protein isoforms, differential protein expressions (7,8,10,11). Limitations and biases of traditional proteomics stunt functional analysis (18).…”

“…Proteomic analysis of spermatogenesis (7) and porcine (8), mouse (9), human testis (10,11) provided useful resources for studying of the testis developmental biology and pathology. However, most of these data were not publicly available and were not validated furtherly on gene or protein levels, and it also included many abundance proteins.…”

Proteomic analysis of reproduction proteins involved in litter size from porcine placenta

“…[14][15][16] Thus far, only a few pig reproduction-associated proteomic studies using reproduction organs such as oocytes, testis, oviductal epithelium, and boar spermatozoa have been conducted. [17][18][19][20] Lee et al 21) reported that placental insufficiency in a cloned pig obtained by somatic cell-derived nuclear transfer showed high relevance to the apoptotic signaling pathway including 14-3-3 and Annexin V proteins compared to normal placenta using two-dimensional gel electrophoresis (2-DE) analysis. Although some of the mechanisms involved in reproduction have been revealed using the 2-DE approach, this method is limited due to poor sensitivity, low throughput, and low reproducibility.…”