Proteomic identification of human sperm proteins

Abstract

Conventional 1-DE has in the past provided a wealth of information concerning the major sperm proteins. However, so far there are relatively few reports exploiting the potential of the present proteomic tools to identify and to study additional yet-unidentified important proteins present in human spermatozoa. In the present work, 2-DE of proteins extracted from human normozoospermic spermatozoa led to the resolution of over 1000 spots. Subsequent excision from the gels of 145 spots and MALDI-TOF MS analysis allowed the identification of 98 different proteins. The function of these proteins turned out to be energy production (23%), transcription, protein synthesis, transport, folding and turnover (23%), cell cycle, apoptosis and oxidative stress (10%), signal transduction (8%), cytoskeleton, flagella and cell movement (10%), cell recognition (7%), metabolism (6%) and unknown function (11%). As many as 23% of the proteins identified have not been previously described as being expressed in human spermatozoa. The present data provide an important clue towards determining the function of these proteins and opens up the possibility to perform additional experiments.

Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker

“…Reverse transcription-polymerase chain reactions (RT-PCR) for the synthesis of complementary DNA (cDNA), 12 μL aliquots of master mixture containing 100 ng of human testes RNA, 1 μL of 500 ng/μL oligo(dT) [12][13][14][15][16][17][18] primer (Invetrogen, Grand Island, NY, USA), and 9 μL of diethylpyrocarbonate-treated water were heated to 70°C for 10 min and put on ice. RT reactions were performed in 20 μL containing master mixture, 4 μL of 5X first-strand synthesis buffer, 0.1 M dithiothreitol, 10 mM of each dNTP, and 200 units of Superscript TM II RNase H -reverse transcriptase (Invetrogen, Grand Island, NY, USA).…”

“…HIBADH is expressed in rat liver, heart, muscle, kidney, and cultural neural cells [34,41]. A larger-scale proteomic study identified approximately 100 proteins in mature human sperm, including HIBADH [18]. However, its detailed expressional patterns in different human organs and developmental stages have not Fig.…”

“…A proteomic approach has been used to identify unknown proteins involved in spermatozoa in differential species [18][19][20]. This approach has been used specifically to discover differential sperm-related proteins (e.g., sperm-surface proteins and antisperm antibodies) [21][22][23].…”

Proteomics of spermatogenesis: from protein lists to understanding the regulation of male fertility and infertility

“…In 2005, Johnston et al 42 used an LC-MS approach to identify 1760 proteins in human sperm. Then in 2006, Martínez-Heredia et al 43 provided a reference two-dimensional map of the mature human sperm proteome with 131 proteins identified. Other studies have also revealed a similar number of proteins.…”

Identification of Guanylate Cyclases and Related Signaling Proteins in Sperm Tail from Sea Stars by Mass Spectrometry

“…ATP synthase also had hits with high scores, which is consistent with the importance of energy production in sperm. Martinez-Heredia et al (2006) identified 98 proteins in human sperm using 2D-gel and matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. We identified 74 proteins in the sea star sperm tail.…”