Proteomic Characterization of Zinc-Binding Proteins of Canine Seminal Plasma

ABSTRACT: Studies have been performed to identify the proteins present in canine seminal plasma (SP) and relate them to sperm quality as well as to discover molecular markers of reproductive tract diseases. There is evidence that heparin-binding proteins, zinc-binding proteins, and lactoferrin as well as the matrix metalloproteinase, superoxide dismutase, catalase, and glutathione peroxidase enzymes are associated with canine sperm quality. Other studies indicate that prolactin and enzymes like arginine esterase, acid phosphatase, and alkaline phosphatase could be successfully used as biomarkers of reproductive disorders. Thus, the present literature review aims to address aspects related to proteins of the canine SP, their influence on fertility, and their importance as biomarkers of reproductive disorders.

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Effect of zinc-binding proteins of seminal plasma on motility and acrosome integrity of canine spermatozoa stored at 5°C

Reproductive Biology ◽

10.1016/j.repbio.2013.01.022 ◽

2013 ◽

Vol 13 ◽

pp. 26-27

Author(s):

M. Mogielnicka-Brzozowska ◽

L. Fraser ◽

A. Dziekońska ◽

K. Tołścik ◽

R. Strzeżek ◽

...

Keyword(s):

Seminal Plasma ◽

Binding Proteins ◽

Zinc Binding ◽

Acrosome Integrity

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Identification and characterization of non-phosphorylcholine-binding and phosphorylcholine-binding proteins of canine seminal plasma

Annals of Animal Science ◽

10.1515/aoas-2017-0005 ◽

2017 ◽

Vol 17 (3) ◽

pp. 771-786 ◽

Cited By ~ 3

Author(s):

Marzena Mogielnicka-Brzozowska ◽

Mariola Słowińska ◽

Leyland Fraser ◽

Paweł Wysocki ◽

Rafał Strzeżek ◽

...

Keyword(s):

Seminal Plasma ◽

Binding Proteins ◽

Membrane Surface ◽

Immunofluorescent Staining ◽

Acrosomal Membrane ◽

Preferential Binding ◽

Sperm Plasma Membrane ◽

Sperm Membrane ◽

Plasma Membrane Surface

Abstract Seminal plasma (SP) proteins participate in the process of fertilization by binding to the sperm membrane, particularly to the phosphorylcholine-containing lipids. This study aimed to identify and characterize non-phosphorylcholine-binding and phosphorylcholine-binding proteins (nPch- BPs and PchBPs, respectively) of canine SP. The nPchBPs and PchBPs were isolated from canine SP by affinity chromatography. Electrophoretic studies revealed that the nPchBPs and PchBPs occurred in their native state as high-molecular-weight aggregates. Immunofluorescent staining showed preferential binding of nPchBPs to the sperm acrosome membrane, whereas PchBPs coating was uniformly distributed on the sperm post-acrosomal membrane, mid-piece and tail regions. Analysis with mass spectrometry confirmed that canine prostate specific esterase (CPSE) is a component of the nPchBPs and PchBPs, which is implicated in key mechanisms of protein-coating on the sperm plasma membrane surface. In addition, proteins of known binding properties such as prostaglandin-H2 D-isomerase and lipocalin-like 1 protein, identified in canine SP, might have a specific role in the fertilization-associated processes.

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Effect of seminal plasma zinc–binding proteins on motility and membrane integrity of canine spermatozoa stored at 5°C

Bulletin of the Veterinary Institute in Pulawy ◽

10.2478/bvip-2014-0025 ◽

2014 ◽

Vol 58 (1) ◽

pp. 163-168 ◽

Cited By ~ 3

Author(s):

Marzena Mogielnicka-Brzozowska ◽

Anna Dziekońska ◽

Rafał Strzeżek ◽

Michał Załęcki ◽

Anna Majewska ◽

...

Keyword(s):

Seminal Plasma ◽

Binding Proteins ◽

Membrane Damage ◽

Membrane Integrity ◽

Control Sample ◽

Incubation Mixture ◽

Zinc Binding ◽

Surface Binding ◽

Positive Effects ◽

Straight Line

AbstractSperm surface binding sites for non-zinc-binding proteins (nZnBPs) and zinc-binding proteins (ZnBPs) were studied by the fluorescence technique with biotin-labelled proteins. The nZnBPs binding pattern was unspecific, no characteristic sites on plasmalemma were found. ZnBPs were attached mainly to the acrosomal region of sperm head and to the sperm flagellum. ZnBPs added to the incubation mixture of the canine spermatozoa allowed the preservation of higher values for total motility, progressive motility, curvilinear line velocity, straight line velocity, and beat cross frequency (P < 0.05), both at time 0 and after 1 h incubation at 5ºC. The addition of nZnBPs to the incubation mixture caused only weak positive effects when compared with control sample (PBS). A higher percentage of canine-ejaculated spermatozoa with intact membranes were observed when ejaculate was incubated with ZnBPs in comparison to control sample stored with PBS (P < 0.05) or nZnBPs (P < 0.05). Spermatozoa diluted with ZnBPs and nZnBPs exhibited a higher percentage of cells with active mitochondria when compared with control, both at time 0 and after 1 h; however, no statistical differences were observed. Our results emphasise the role of seminal plasma protein in securing the correct quantity and availability of zinc ions as a component regulating the motility of canine spermatozoa. The protective effect of ZnBPs against the cooling effect may be due to their ability of preventing sperm membrane damage.

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Zinc-binding proteins from boar seminal plasma -- isolation, biochemical characteristics and influence on spermatozoa stored at 4°C.

Acta Biochimica Polonica ◽

10.18388/abp.2011_2261 ◽

2011 ◽

Vol 58 (2) ◽

Cited By ~ 10

Author(s):

Marzena Mogielnicka-Brzozowska ◽

Paweł Wysocki ◽

Jerzy Strzeżek ◽

Władysław Kordan

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Binding Proteins ◽

Control Sample ◽

Zinc Binding ◽

Shielding Effect ◽

Sperm Plasma Membrane ◽

Native Electrophoresis ◽

Seminal Plasma Proteins ◽

Fast Flow

Affinity chromatography on Chelating Sepharose Fast Flow Gel-Zn(2+) was used for fractionation of boar seminal plasma proteins. Approximately 30% of total boar seminal plasma proteins showed affinity for zinc ions (ZnBP fraction). Native electrophoresis (PAGE) of ZnBP revealed six protein fractions which separated into 27 bands under denaturing conditions (SDS/PAGE). Two-dimensional electrophoresis (2D PAGE) showed 148 polypeptides with isoelectric points mostly in the basic and neutral pH range. The zinc-binding proteins comprise mainly 10-20 kDa polypeptides which are probably members of the spermadhesin family. ZnBP present in the incubation mixture of spermatozoa stored for 1 or 24 h at 4 °C allowed preservation of a higher percentage of cells exhibiting linear motility in comparison to a control sample stored in PBS. Presented results indicate that proteins binding Zn(2+) ions have a shielding effect on the sperm plasma membrane and acrosome of spermatozoa, protecting these structures against consequences of cold shock.

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