Aggregated Forms of Bull Seminal Plasma Proteins and Their Heparin-Binding Activity

2004 ◽  
Vol 69 (3) ◽  
pp. 616-630 ◽  
Author(s):  
Petra Jelínková ◽  
Helena Ryšlavá ◽  
Jiří Liberda ◽  
Věra Jonáková ◽  
Marie Tichá
Keyword(s):  
Molecular Weight ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Sodium Dodecyl ◽  
Reversed Phase ◽  
Binding Activity ◽  
Size Exclusion ◽  
Heparin Binding ◽  
Aggregation State ◽  
Seminal Plasma Proteins

Heparin-binding activity of bull seminal plasma proteins was shown to be dependent on their aggregation state. The protein fraction interacting with immobilized heparin was characterized by large polydispersity in the region of molecular weight of 60 000-10 000, while that not retained on the affinity carrier was present as aggregates with molecular weight >100 000. Components of heparin-binding and non-heparin-binding fractions were separated by RP HPLC (reversed-phase HPLC) and analyzed by SDS (sodium dodecyl sulfate) electrophoresis and N-terminal sequencing. Size exclusion chromatography of whole seminal plasma and heparin-binding proteins in the presence of D-fructose (as a component of seminal plasma) showed that the region of molecular weights of protein-associated forms was shifted to lower values. An increase of heparin-binding activity of bull proteins, as determined by ELBA (Enzyme-Linked Binding Assay), correlates with a decrease of their aggregation state. The modulation of the aggregation state of bull proteins by seminal plasma components and, in this way, also of their heparin-binding properties suggests possible mechanisms for capacitation mediated by these proteins.

scholarly journals Amino acid sequence of HSP-1, a major protein of stallion seminal plasma: effect of glycosylation on its heparin- and gelatin-binding capabilities

1995 ◽  
Vol 310 (2) ◽  
pp. 615-622 ◽  
Author(s):  
J J Calvete ◽  
K Mann ◽  
W Schäfer ◽  
L Sanz ◽  
M Reinert ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Seminal Plasma ◽  
Binding Activity ◽  
Size Exclusion ◽  
Major Protein ◽  
Heparin Binding ◽  
Aggregation State ◽  
Consensus Pattern

We report the complete amino acid sequence of HSP-1, a major protein isolated from stallion seminal plasma or acid extracts of ejaculated spermatozoa. The protein consists of 121 amino acids organized in two types of homologous repeats arranged in the pattern AA‘BB’. Each of the 13-15-residue A-type repeats contains two O-linked oligosaccharide chains. The B-type repeats span 44-47 amino acids each, are not glycosylated, and have the consensus pattern of the gelatin-binding fibronectin type-II module. This domain also occurs in the major bovine seminal plasma heparin-binding proteins PDC-109 (BSP-A1/A2) and BSP-A3. However, unlike the bovine proteins which bind quantitatively to a heparin-Sepharose column, stallion HSP-1 was recovered in both the flow-through and the heparin-bound fractions. Structural analysis showed that the two HSP-1 forms contain identical polypeptide chains which are differently glycosylated. Moreover, size-exclusion chromatography showed that heparin-bound HSP-1 associates with HSP-2, another major seminal plasma protein, into a 90 kDa product, whereas the non-heparin-bound glycoform of HSP-1 is eluted as a monomeric (14 kDa) protein. This suggests that glycosylation may have an indirect effect on the heparin-binding ability of HSP-1 through modulation of its aggregation state. On the other hand, both glycoforms of HSP-1 displayed gelatin-binding activity, indicating that the molecular determinants for binding heparin and gelatin are different.

Seminal plasma proteins inhibit in vitro- and cooling-induced capacitation in boar spermatozoa

2010 ◽  
Vol 22 (6) ◽  
pp. 893 ◽  
Author(s):  
Melissa L. Vadnais ◽  
Kenneth P. Roberts
Keyword(s):  
Mass Spectrometry ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Acrosome Reaction ◽  
Heparin Binding ◽  
Total Proteins ◽  
Binding Properties ◽  
Seminal Plasma Proteins ◽  
Boar Spermatozoa

Dilute boar seminal plasma (SP) has been shown to inhibit in vitro capacitation and cooling-induced capacitation-like changes in boar spermatozoa, as assessed by the ability of the spermatozoa to undergo an ionophore-induced acrosome reaction. We hypothesised that the protein component of SP is responsible for this effect. To test this hypothesis, varying concentrations of total SP protein or SP proteins fractionated by heparin binding were assayed for their ability to inhibit in vitro capacitation, as well as cooling- and cryopreservation-induced capacitation-like changes. In vitro capacitation and cooling-induced capacitation-like changes were prevented by 10% whole SP, as well as by total proteins extracted from SP at concentrations greater than 500 μg mL−1. No amount of SP protein was able to prevent cryopreservation-induced capacitation-like changes. Total SP proteins were fractionated based on their heparin-binding properties and the heparin-binding fraction was shown to possess capacitation inhibitory activity at concentrations as low as 250 µg mL−1. The proteins in the heparin-binding fraction were subjected to mass spectrometry and identified. The predominant proteins were three members of the spermadhesin families, namely AQN-3, AQN-1 and AWN, and SP protein pB1. We conclude that one or more of these heparin-binding SP proteins is able to inhibit in vitro capacitation and cooling-induced capacitation-like changes, but not cryopreservation-induced capacitation-like changes, in boar spermatozoa.

scholarly journals Electrophoretic profile of seminal proteins and their correlation with in vitro sperm characters in Black Bengal buck semen

2019 ◽  
Vol 12 (5) ◽  
pp. 621-628 ◽  
Author(s):  
M. Karunakaran ◽  
Vivek C. Gajare ◽  
Ajoy Mandal ◽  
Mohan Mondal ◽  
S. K. Das ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Membrane Integrity ◽  
Sperm Proteins ◽  
Sds Page ◽  
Sperm Characters ◽  
Functional Membrane ◽  
Seminal Plasma Proteins

Aim: This study aimed to study the electrophoretic properties of seminal plasma and sperm proteins of Black Bengal buck semen and their correlation with in vitro sperm characters and freezability. Materials and Methods: Semen ejaculates from nine Black Bengal bucks were collected by artificial vagina (n=20/buck). Ejaculates were evaluated for in vitro sperm characters and electrophoretic profile of seminal protein. In vitro sperm characters were evaluated immediately after collection, after completion of equilibration period, and after freeze-thawing. For seminal protein studies, seminal plasma proteins were precipitated by ice-cold ethanol method, and sperm proteins were extracted by Triton X detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to assess the molecular weight of seminal proteins. Correlation between in vitro sperm characters and protein bands was determined by Pearson's correlation coefficient, and two-way ANOVA was applied to find the individual buck differences. Results: Significant difference (p<0.01) among the bucks was noticed in the in vitro sperm characters evaluated at all the three stages of semen evaluation such as immediately after collection, after completion of equilibration period, and post-freeze thawing. Progressive loss of sperm motility, membrane integrity, and other in vitro sperm characters were noticed during cryopreservation. A total of ten protein bands in the molecular weight ranging from 17 to 180 kDa were found in the SDS-PAGE of seminal plasma proteins, while nine bands of 17-134 kDa were observed in sperm proteins. Seminal plasma proteins of molecular weight 75, 62-49, 20, and 17 kDa and sperm proteins of 75, 20, and 17 kDa were present in all the nine bucks (100%) screened, and variation among the bucks was noticed for the presence of other proteins. Seminal plasma protein of 180-134 kDa showed a negative correlation with individual motility (−0.716) and functional membrane integrity of sperm cells (−0.724) in post-freeze-thaw analysis and 48 kDa protein had a positive correlation with individual motility (0.649) and functional membrane integrity of sperm cells (0.664) in post-thaw analysis. Sperm proteins of 63 kDa had a negative correlation (−0.616) with sperm concentration in neat semen. Conclusion: Variation among the bucks was noticed in the in vitro sperm characters and semen freezability. Correlation between seminal proteins and in vitro sperm characters and semen freezability had been found which might be useful as a tool to select breeding bucks.

Effect of glycosylation on the heparin-binding capability of boar and stallion seminal plasma proteins

1995 ◽  
Vol 711 (1) ◽  
pp. 167-173 ◽  
Author(s):  
Juan J. Calvete ◽  
Markus Reinert ◽  
Libia Sanz ◽  
Edda Töpfer-Petersen
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Heparin Binding ◽  
Seminal Plasma Proteins ◽  
Binding Capability

Functional attributes of seminal proteins in bull fertility: A systematic review

Reproduction ◽  
2021 ◽  
Author(s):  
Arabela Guedes de Azevedo Viana ◽  
Iara Magalhães Ribeiro ◽  
Renner Philipe Rodrigues Carvalho ◽  
Erdogan Memili ◽  
Arlindo Alencar Moura ◽  
...  
Keyword(s):  
Systematic Review ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Protein Isoforms ◽  
Heparin Binding ◽  
Protein Biomarkers ◽  
Functional Attributes ◽  
Fertilizing Ability ◽  
Seminal Plasma Proteins ◽  
Sperm Fertilizing Ability

Proteomic approaches have been widely used in reproductive studies to uncover protein biomarkers of bull fertility. Seminal plasma is one of the most relevant sources of these proteins that may influence sperm physiology. Nonetheless, there are still gaps in existing knowledge in the functional attributes of seminal proteins. Thus, we reviewed the relationships between seminal plasma proteins and bull fertility by conducting a systematic review with data obtained from 71 studies. This review showed that the associations between fertility improvement with the use of total seminal plasma proteins are still controversial. None of the studies explored the sperm fertilizing ability following these interactions. By contrast, the exposure to a single protein, such as osteopontin, binder of sperm proteins, and heparin binding proteins, can increment sperm motility, capacitation, and fertilizing ability by modulating intracellular calcium concentrations, removing lipids from sperm membranes, and regulating the acrosome reaction. Variations in protein analyses and the protein contents and their abundances between animals contributed to the difficulty of establishing protein biomarkers of fertilizing potential of the bull sperm. Indeed, the heterogenicity of methodologies was a limitation of this review. Standardized methods of seminal protein analyses, as well as sperm endpoints, may minimize such discrepancies. In conclusion, potential biomarkers of sperm parameters are still to be established. Future studies should evaluate protein isoforms and how they interact with sperm to ascertain their biological functions.

scholarly journals Reproductive biotechnology in animal husbandry - current status and future prospects

2015 ◽  
Vol 31 (4) ◽  
pp. 467-480 ◽  
Author(s):  
M. Ivanova ◽  
D. Gradinarska ◽  
D. Daskalova
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Animal Husbandry ◽  
Biological Evaluation ◽  
Size Exclusion ◽  
Current Status ◽  
Computer Assisted ◽  
Biological Potential ◽  
Seminal Plasma Proteins

To date there is still no optimal biotechnology which ensures maximum preservation of the functional parameters of the spermatozoa from buffaloes, boars and dogs. The aim of this research is to study the biological potential of seminal plasma proteins that are specific only to ejaculates with high cryotolerance and good quality parameters of the spermatozoa. The motility and velocity parameters of the spermatozoa were assessed by computer-assisted sperm analysis. Seminal plasma proteins were separated by size-exclusion liquid chromatography and characterized by polyacrylamide gel electrophoresis and mass spectrometry. Based on the results obtained, sperm diluents and methods for biological evaluation of the fertilization potential of the spermatozoa from buffalo bulls, boars and dogs were created and proposed for practical application.

Characteristics of selected seminal plasma proteins and their application in the improvement of the reproductive processes in mammals

2011 ◽  
Vol 14 (3) ◽  
pp. 489-499 ◽  
Author(s):  
M. Mogielnicka-Brzozowska ◽  
W. Kordan
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Chromatin Condensation ◽  
Female Reproductive Tract ◽  
Sperm Chromatin ◽  
Heparin Binding ◽  
Seminal Plasma Proteins ◽  
Reproductive Processes

Characteristics of selected seminal plasma proteins and their application in the improvement of the reproductive processes in mammals Understanding the biochemical processes associated with ovum fertilization and knowledge about the structure and function of individual substances participating in these processes is crucial for the development of biotechnological methods to improve reproduction of animals and humans. Among many components of seminal plasma, proteins and peptides play a specific role in regulation of the fertilization process, particularly through their ability to bind various types of ligands such as polysaccharides, lipids and ions. Heparin-binding proteins regulate capacitation and acrosome reaction processes. Affinity of plasma proteins to mannans of the fallopian tube epithelium facilitates formation of spermatozoa reservoirs in the female reproductive tract. Ability to bind phosphorylcholine is one of the conditions for the coating of the seminal plasma proteins on the sperm membrane and also determines the formation of oligomeric forms of certain proteins. Zinc binding by seminal plasma proteins regulates sperm chromatin condensation state. It also affects motility of these cells and acrosome reaction. The interspecies analysis indicates significant structural and functional similarities, especially for the proteins with low molecular weight. Fertility associated proteins (FAPs) have been determined in the bull, stallion, boar, ram and dog. The contents of these proteins correlate with the indicators of the fertilizing abilities of sperm. In humans, several seminal plasma proteins were found which serve as diagnostic markers of spermatogenesis, seminiferous epithelium state, and azoospermia. To determine the semen ability for preservation, measurement of some seminal plasma protein content may also be used. Addition of specific plasma proteins to a spermatozoa solution undergoing the process of preservation may be used to retain the features of the cells responsible for efficient fertilization.

Comparison of the Non-Specific Binding of Unfractionated Heparin and Low Molecular Weight Heparin (Enoxaparin) to Plasma Proteins

1993 ◽  
Vol 70 (04) ◽  
pp. 625-630 ◽  
Author(s):  
Edward Young ◽  
Benilde Cosmi ◽  
Jeffrey Weitz ◽  
Jack Hirsh
Keyword(s):  
Molecular Weight ◽  
Unfractionated Heparin ◽  
Plasma Proteins ◽  
Low Molecular Weight Heparin ◽  
Specific Binding ◽  
Anticoagulant Activity ◽  
Factor Xa ◽  
Low Molecular Weight ◽  
Heparin Binding ◽  
Fixed Amount

SummaryThe non-specific binding of anticoagulantly-active heparin to plasma proteins may influence its anticoagulant effect. We used low affinity heparin (LAH) essentially devoid of anti-factor Xa activity to investigate the extent and possible mechanism of this non-specific binding. The addition of excess LAH to platelet-poor plasma containing a fixed amount of unfractionated heparin doubled the anti-factor Xa activity presumably because it displaces anticoagulantly-active heparin from plasma proteins. Although dextran sulfates of varying molecular weights also increased the anti-factor Xa activity, less sulfated heparin-like polysaccharides had no effect. These findings suggest that the ability to displace active heparin from plasma protein binding sites is related to charge and may be independent of molecular size. In contrast to its effect in plasma containing unfractionated heparin, there was little augmentation in anti-factor Xa activity when LAH was added to plasma containing low molecular weight heparin (LMWH), indicating that LMWH binds less to plasma proteins than unfractionated heparin. This concept is supported by studies comparing the anticoagulant activity of unfractionated heparin and LMWH in plasma with that in buffer containing antithrombin III. The anti-factor Xa activity of unfractionated heparin was 2-fold less in plasma than in the purified system. In contrast, LMWH had identical anti-factor Xa activity in both plasma and buffer, respectively. These findings may be clinically relevant because the recovered anti-factor Xa activity of unfractionated heparin was 33% lower in plasma from patients with suspected venous thrombosis than in plasma from healthy volunteers. The reduced heparin recovery in patient plasma reflects increased heparin binding to plasma proteins because the addition of LAH augmented the anti-factor Xa activity. In contrast to unfractionated heparin, there was complete recovery of LMWH added to patient plasma and little increase of anti-factor Xa activity after the addition of LAH. These findings may explain why LMWH gives a more predictable dose response than unfractionated heparin.

Genetic Polymorphism of Boar Seminal Plasma Proteins

1993 ◽  
Vol 64 (3) ◽  
pp. 221-227
Author(s):  
Soichi TSUJI ◽  
Masatoshi ASAO ◽  
Hiroshi KUSUNOKI ◽  
Takao OISHI
Keyword(s):  
Genetic Polymorphism ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Seminal Plasma Proteins
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