Proteomic analysis of sperm proteins in infertile men with high levels of reactive oxygen species

Abstract

Oxidative stress is a significant risk factor for male infertility. A pro-oxidant testicular environment may alter the expression profile of functional sperm proteins and result in poor sperm quality. Patients and donors were divided into ROS (-) and ROS (+) groups. Using computational studies, and data mining of available literature on spermatozoa, oxidative stress and proteomics, we identified three core regulatory proteins angiotensin-converting enzyme (ACE), heat-shock protein (Hsp70) family A member 2 (HSPA2) and ribosomal protein subunit 27A (RPS27A) and seven interlink proteins NOS2, SUMO2, UBL4A, FBXO25, MAP3K3, APP and UBC. HSPA2 was validated by Western Blot, while the localisation of ACE, RPS27A, MAP3K3 and APP was identified by immunocytochemistry. The obtained results showed that HSPA2 was 1.2 (ROS+) and 2.1 (ROS-) fold downregulated in spermatozoa from patients with high levels of reactive oxygen species (ROS). ACE and APP were localised in the post-acrosomal region of spermatozoa, whereas RPS27A and MAP3K3 were localised either in the tail or sperm neck area. Our data show that these proteins may play a role in ROS-induced male infertility.

Alterations of Spermatozoa Proteomic Profile in Men with Hodgkin's Disease Prior to Cancer Therapy

“…ACE is a protein involved in fertility and our IPA analysis relates this protein to binding of sperm and zona pellucida, and interrelates the binding of sperm with fertilization process and fertility. ACE has been reported to be altered in infertile patients [ 13 27 28 ]. In our experiment, the proteomic analysis showed an overexpression in ACE, although the WB validation did not show any significant increase in expression.…”

“…Due to the localization of ACE in the peri-acrosomal section of the sperm head, and an agonist-induced function in acrosomal exocytosis, this protein consequently has a significant role in the fertility process [ 30 ]. Studies have reported a correlation between decrease in ACE with an increase in ROS and possible impairment of acrosome reaction [ 28 29 ]. PRDXs are central antioxidant enzymes and have a role in sperm function and male fertility [ 31 ].…”

Molecular Changes Induced by Oxidative Stress that Impair Human Sperm Motility

“…Similarly, reduced levels of AKAP4 have also been implicated as a biomarker of human asthenozoospermic males [95]. In addition to AKAP4, alternative cAMP-responsive elements such as protein kinase A (PKA), have also been validated as primary targets of 4HNE-mediated modification [22], whilst seminal OS has been correlated with altered levels of several proteins mapping to the MAPK/ERK pathways [123]. Such defects have been correlated with a commensurate decrease in global levels of tyrosine phosphorylation in human spermatozoa [22]; a form of post-translational modification that underpins sperm motility, and is particularly important in the induction of hyperactivation [124].…”

Proteomic Analyses of Human Sperm Cells: Understanding the Role of Proteins and Molecular Pathways Affecting Male Reproductive Health

“…Under oxidative stress condition, proteins associated with carbohydrate metabolic pathways, such as gluconeogenesis and glycolysis, and protein modification are compromised in sperm cells [112]. Ayaz et al demonstrated that proteins related to sperm function such as acrosome reaction and flagellar movement (ACE, HSPA2, RPS27A, MAP3K3 and APP) are altered in the ROS-induced male infertility [113]. Furthermore, stress response and cellular, metabolic and regulatory pathways are dysregulated in the sperm of infertile men with increased levels of ROS [50].…”