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

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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Proteins and Heparin Binding Proteins of Spermatozoa and Seminal Plasma in Beetal Bucks: Purification, Characterization and Role in In vivo Fertility

Indian Journal of Animal Research ◽

10.18805/ijar.b-3974 ◽

2020 ◽

Author(s):

Ranjna S. Cheema ◽

Navjot S. Dhillon ◽

Sumit Singhal

Keyword(s):

Seminal Plasma ◽

Binding Proteins ◽

Proteome Analysis ◽

Fertility Rate ◽

Heparin Binding ◽

Sds Page ◽

Sperm Extract ◽

Sperm Membrane ◽

Special Relevance

Background: The proteome analysis of seminal plasma and spermatozoa is of special relevance in livestock. Heparin binding proteins (HBPs) found in the seminal plasma of several mammals are shown to bind to sperm membrane and affect a series of events that contribute to normal fertility, such as sperm capacitation, formation of the oviduct reservoir and binding to the oocyte. Profiles of HBPs from seminal plasma and sperm membranes have been associated with sperm fertility. Although, HBPs present in the SP are described in several species, but little is known about HBPs in buck. Methods: Seminal plasma (SP) and sperm extracts (SE) of 13 bucks were subjected to heparin-sepharose affinity chromatography. Sperm extract, seminal plasma and purified HBPs and Non-HBPs were fractionated by SDS-PAGE. Total 78 females (6 per buck) were mated with 13 bucks. Bucks were divided into two groups, G-I (high fertile, 83.3-100% FR) and G-II (low fertile, 50-66.7% FR). Relationship between HBPs and fertility rate was observed. Result: SDS – PAGE of SP and SE resulted in resolution of 22 (10-240 kDa) and 21 (10-270 kDa) bands, respectively. Based on fertility rate 15 and 13 kDa proteins were absent in SP of higher number of GI-compared to G-II bucks. Fourteen bands ranging from 10 – 180 kDa and 10 – 150 kDa were separated from SP-NHBP and SP-HBP. SP-HBPs of 75, 35, 30, 28, 25 and 13 kDa were present in higher (28.6%, 42.5%, 26.2%, 40.5%, 14.3% and 36.2%) number of high fertile than low fertile bucks. NHBP and HBP purified from SE resolved into 11 bands ranging from 10 – 135 kDa and 10 – 120 kDa, respectively. SE-HBP of 53 kDa, 50/45 kDa and 25 kDa were present in higher percentage of high fertile than low fertile bucks.

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Proteomic Characterization of Zinc-Binding Proteins of Canine Seminal Plasma

Reproduction in Domestic Animals ◽

10.1111/rda.12629 ◽

2015 ◽

Vol 50 (6) ◽

pp. 1017-1021 ◽

Cited By ~ 7

Author(s):

M Mogielnicka-Brzozowska ◽

N Kowalska ◽

L Fraser ◽

W Kordan

Keyword(s):

Seminal Plasma ◽

Binding Proteins ◽

Zinc Binding

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Characterization of Tumor Specific Antigens on the Plasma Membrane Surface of Rat Hepatomas lnduced by 3'-Me DAB and ldentification of the Common Tumor Specific Antigens from Rat Hepatomas lnduced by Different Chemical Hepatocarcinogens

Yonsei Medical Journal ◽

10.3349/ymj.1988.29.1.17 ◽

1988 ◽

Vol 29 (1) ◽

pp. 17

Author(s):

Yoon Soo Kim ◽

Kyung-Soo Hahm ◽

Kyung Sup Kim ◽

Nam Jeen Lee

Keyword(s):

Plasma Membrane ◽

Membrane Surface ◽

Specific Antigens ◽

The Common ◽

Plasma Membrane Surface

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Role of seminal plasma phospholipid-binding proteins in sperm membrane lipid modification that occurs during capacitation

Journal of Reproductive Immunology ◽

10.1016/s0165-0378(01)00098-5 ◽

2002 ◽

Vol 53 (1-2) ◽

pp. 109-119 ◽

Cited By ~ 175

Author(s):

Puttaswamy Manjunath ◽

Isabelle Thérien

Keyword(s):

Seminal Plasma ◽

Binding Proteins ◽

Plasma Phospholipid ◽

Membrane Lipid ◽

Lipid Modification ◽

Phospholipid Binding ◽

Sperm Membrane

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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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