Proteomic Analysis of Seminal Plasma from Normal Volunteers and Post-Vasectomy Patients Identifies over 2000 Proteins and Candidate Biomarkers of the Urogenital System

Abstract

Seminal plasma is a fluid that originates from the testis, epididymis,prostate, and seminal vesicles, and hence, proteomic studies may identify potential markers of infertility and other diseases of the genito-urinary tract. We profiled the proteomes of pooled seminal plasma from fertile Control and post-vasectomy (PV) men. PV seminal plasma samples are void of proteins originating from the testis and the epididymis due to ligation of the vas deferens, and hence, comparative analysis of Control and PV data sets allows for identification of proteins originating from these tissues. Utilizing offline MudPIT and high-resolution mass spectrometry, we were able to identify over 2000 proteins in Control and PV pools each and over 2300 proteins all together. With semiquantitative analysis using spectral counting, we catalogued 32 proteins unique to Control, 49 at lower abundance in PV, 3 unique to PV, and 25 at higher abundance in PV. We believe that proteins unique to Control or at lower abundance in PV have their origin in the testis and the epididymis. Public databases have confirmed that many of these proteins originate from the testis and epididymis and are linked to the reproductive tract. These proteins may serve as candidate biomarkers for future studies of infertility and urogenital diseases.

Proteomics of human seminal plasma: Identification of biomarker candidates for fertility and infertility and the evolution of technology

“…In fact, most of these proteins, such as prostatic-specific antigen (PSA), semenogelin I, and semenogelin II, are involved in the regulation, processing, or degradation of seminal fluid proteins and the coagulation of semen. This large number of enzymes is consistent with the statement that a great number of seminal proteins are involved in semen coagulum liquefaction and sperm capacitation (Pilch and Mann, 2006;Batruch et al, 2011).…”

“…The flight time of each ionized fragment is proportional to the square root of the m/z ratio. As the charge (z) of the ionized proteins is often þ1, the m/z value is often equal to the mass (2003) MALDI-TOF Impaired spermatogenesis Utleg et al (2003) LCMS/MS Prostasome Fung et al (2004) MALDI-TOF; LC-MS/MS Healthy fertile men Pilch and Mann (2006) MALDI-TOF; LTQ-FT Healthy fertile man Yang et al (2007) SELDI-TOF Biomarkers of oligospermia Yamakawa et al (2007) LCMS/MS Infertile versus fertile men Bai et al (2008) SELDI-TOF Severe oligospermia versus healthy fertile men Kumar et al (2008) MALDI-TOF Heparin binding proteins in human seminal plasma Kumar et al (2009) MALDI-TOF Heparin binding proteins in human seminal plasma Wang et al (2009) LCMS/MS Asthenozoospermia Drake et al (2010) LTQ-Orbitrap XL Prostatic secretions Batruch et al (2011) LTQ-Orbitrap XL Post-vasectomy versus healthy fertile men Milardi et al (2012b) LTQ-Orbitrap XL Healthy fertile men Batruch et al (2012) LTQ-Orbitrap XL Non-obstructive azoospermia Kagedan et al (2012) LTQ-Orbitrap XL Prostatitis Molecular Reproduction & Development value. The spectral output produced by MALDI-TOF is represented by a number of protein peaks, which are described by a m/z value on the horizontal axis and by a peak-intensity value on the vertical axis (Schuchardt and Sickmann, 2007).…”

Unraveling the sperm proteome and post-genomic pathways associated with sperm nuclear DNA fragmentation

“…AKAP4 is present in the fibrous sheath of sperm flagellum and is involved with sperm motility and morphology and in GPDHS binding to fibrous sheath [57]. SPACA3 is an acrosome protein and is important to sperm-egg fusion during fertilization [58].…”

“…SLC2A14 is a glucose transmembrane transporter that may act specifically during spermatogenesis, being important for glucose intracellular metabolism [80]. PGK2 is important for glycolysis and was observed in the seminal plasma from fertile men by Batruch et al [58].…”

Identification of biochemical differences between different forms of male infertility by nuclear magnetic resonance (NMR) spectroscopy

“…Seminal plasma is the liquid component of semen and composed of secretions from the testis, epididymis and male accessory glands [6]. At pH 7.35-7.50, it has buffering properties, protecting spermatozoa from the acidic environment of the vagina.…”