Novel purification method for mammalian seminal plasma phospholipid-binding proteins reveals the presence of a novel member of this family of protein in stallion seminal fluid.
Abstract: A family of bull seminal plasma (BSP) phospholipid-binding proteins (BSP proteins), potentiate heparin- and HDL-induced capacitation. The homologous proteins have been purified from stallion and boar seminal plasma, and detected in low concentrations in other mammalian seminal plasma. In this study, we developed a new isolation method for mammalian seminal plasma choline phospholipid-binding proteins wherein they are present in low concentrations. The method is based on the interaction of this family of proteins with egg yolk low-density lipoprotein fraction (LDF). In order to demonstrate the feasibility of the method, we incubated LDF with alcohol precipitates of bull, boar, and stallion seminal plasma. LDF were re-isolated by ultracentrifugation along with bound proteins. LDF with associated proteins were dialyzed, lyophilized, and delipidated. BSP homologous proteins were finally purified by p-aminophenyl phosphorylcholine (PPC)-agarose and/or gelatin-agarose chromatographies, and analyzed by SDS-PAGE. With this new protocol, phospholipid-binding proteins of bull, boar, and stallion seminal plasma were recovered almost 100%. A new 12 kDa stallion seminal plasma protein of the same family was also isolated and partially sequenced. The radio-immunoassay (RIA) data showed that 10 mg of LDF can bind all BSP proteins present in 120 mg of alcohol precipitated BSP proteins. These results confirm the efficiency of the method and that the LDF step could be used for the isolation of all BSP proteins homologs from different mammalian species.
Copyright 2003 Wiley-Liss, Inc.
Publication Date: 2003-10-28 PubMed ID: 14579411DOI: 10.1002/mrd.10369Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research revolves around the development of a novel method for purifying phospholipid-binding proteins found in mammalian seminal fluid, which also revealed the presence of a new member of this protein type in stallion seminal fluid.
Method Development
- The researchers devised a new isolation method for purifying choline phospholipid-binding proteins which are a part of mammalian seminal plasma and are often found in low concentrations.
- This method relies on the interaction of these proteins with the egg yolk’s low-density lipoprotein fraction (LDF).
- As a part of the study, the team incubated LDF with alcohol precipitates from the seminal plasma of bulls, boars, and stallions.
- The LDF, along with bound proteins, were successfully re-isolated through ultracentrifugation.
- Post this, the LDF and associated proteins underwent dialysis, lyophilization, and delipidation.
- Finally, the homologous proteins of bull seminal plasma (BSP) were purified through p-aminophenyl phosphorylcholine (PPC)-agarose or gelatin-agarose chromatographies.
- The purified proteins were then scrutinized through SDS-PAGE, a common laboratory technique in biochemistry for the separation of proteins based on their electrophoretic mobility.
Recovery and Detection of New Protein
- Using this newly devised protocol, the team was able to recover nearly 100% phospholipid-binding proteins present in the seminal plasma of bulls, boars, and stallions.
- In the course of the study, the researchers also discovered and partially sequenced a novel 12 kDa protein in stallion seminal plasma which falls in the same family.
- The radioimmunoassay (RIA) data collected suggested that 10 mg of LDF is capable of binding all BSP proteins present in 120 mg of alcohol precipitated BSP proteins.
Significance of the Study
- The study’s findings improved the process of phospholipid-binding proteins isolation and confirmed the effectiveness of this new method.
- Moreover, the LDF step in the process proved to be beneficial for isolating all homologs of BSP proteins from various mammalian species.
Cite This Article
APA
Ménard M, Nauc V, Lazure C, Vaillancourt D, Manjunath P.
(2003).
Novel purification method for mammalian seminal plasma phospholipid-binding proteins reveals the presence of a novel member of this family of protein in stallion seminal fluid.
Mol Reprod Dev, 66(4), 349-357.
https://doi.org/10.1002/mrd.10369 Publication
Researcher Affiliations
- Department of Medicine and of Biochemistry, University of Montreal, Q, Canada.
MeSH Terms
- Amino Acid Sequence
- Animals
- Cattle
- Chromatography, Affinity
- Horses
- Male
- Molecular Sequence Data
- Phospholipids / metabolism
- Protein Binding
- Semen / chemistry
- Seminal Plasma Proteins / chemistry
- Seminal Plasma Proteins / isolation & purification
- Seminal Plasma Proteins / metabolism
- Swine
Citations
This article has been cited 4 times.- El Kadili S, Kirschvink N, Raes M, Bister JL, Archa B, Douaik A, Chentouf M. Influence of Season and Liquid Storage at 16 °C on Beni Arouss Bucks' Semen Quality.. Animals (Basel) 2020 Oct 29;10(11).
- Plante G, Lusignan MF, Lafleur M, Manjunath P. Interaction of milk proteins and Binder of Sperm (BSP) proteins from boar, stallion and ram semen.. Reprod Biol Endocrinol 2015 Aug 15;13:92.
- Jois PS, Plante G, Thérien I, Manjunath P. Functional characterization of the domains of the bovine binder of SPerm 5 (BSP5) protein.. Reprod Biol Endocrinol 2015 Jun 19;13:64.
- Manjunath P, Lefebvre J, Jois PS, Fan J, Wright MW. New nomenclature for mammalian BSP genes.. Biol Reprod 2009 Mar;80(3):394-7.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
