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The Biochemical journal1995; 310 ( Pt 2)(Pt 2); 615-622; doi: 10.1042/bj3100615

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

Abstract: 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.
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Publication Date: 1995-09-01 PubMed ID: 7654203PubMed Central: PMC1135940DOI: 10.1042/bj3100615Google Scholar: Lookup
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  • Comparative Study
  • 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 article focuses on the amino acid sequence of HSP-1, a major protein present in the seminal plasma of stallions, and investigates the effect of its glycosylation on its ability to bind to heparin and gelatin. The researchers concluded that the glycosylation process influences HSP-1’s heparin-binding capability indirectly by modulating its aggregation state while it doesn’t affect gelatin-binding activity.

The Protein HSP-1 and its Characteristics

  • The research delineates the complete amino acid sequence of HSP-1, a principal protein isolated from stallion seminal plasma, or acid extracts of ejaculated spermatozoa.
  • The protein comprises 121 amino acids arranged in two types of homologous repeats, patterned AA’BB’.
  • A-type repeats contain two O-linked oligosaccharide chains and encompass 13-15 amino acids each. In contrast, B-type repeats span 44-47 amino acids each and are not glycosylated.
  • The gelatin-binding fibronectin type-II module is the consensus pattern of B-type repeats. This domain also exists in major bovine seminal plasma heparin-binding proteins PDC-109 (BSP-A1/A2) and BSP-A3.

HSP-1 Glycosylation and its Binding Capabilities

  • HSP-1’s recovery from a heparin-Sepharose column differs from bovine proteins as it was found in both the flow-through and the heparin-bound fractions.
  • Structural analysis indicated the presence of identical polypeptide chains, differently glycosylated in the two HSP-1 forms. This implies that glycosylation might indirectly affect HSP-1’s ability to bind heparin by altering the protein’s aggregation state.
  • Also, the non-heparin-bound glycoform associates with another principal seminal plasma protein, HSP-2, into a 90 kDa product, according to size-exclusion chromatography. In contrast, the non-heparin-bound glycoform of HSP-1 exists as a monomeric (14 kDa) protein.
  • Finally, both glycoforms of HSP-1 displayed gelatin-binding activity demonstrating that the molecular mechanisms for binding heparin and gelatin differ. This suggests that glycosylation might not directly influence the HSP-1’s gelatin-binding activity.

Cite This Article

APA
Calvete JJ, Mann K, Schäfer W, Sanz L, Reinert M, Nessau S, Raida M, Töpfer-Petersen E. (1995). Amino acid sequence of HSP-1, a major protein of stallion seminal plasma: effect of glycosylation on its heparin- and gelatin-binding capabilities. Biochem J, 310 ( Pt 2)(Pt 2), 615-622. https://doi.org/10.1042/bj3100615

Publication

The Biochemical journal
ISSN: 0264-6021
NlmUniqueID: 2984726R
Country: England
Language: English
Volume: 310 ( Pt 2)
Issue: Pt 2
Pages: 615-622

Researcher Affiliations

Calvete, J J
  • Institut für Reproduktionsmedizin, Tierärztliche Hochschule, Hannover-Kirchrode, Germany.
Mann, K
    Schäfer, W
      Sanz, L
        Reinert, M
          Nessau, S
            Raida, M
              Töpfer-Petersen, E

                MeSH Terms

                • Amino Acid Sequence
                • Animals
                • Binding Sites
                • Carbohydrate Conformation
                • Carbohydrate Sequence
                • Carrier Proteins / chemistry
                • Carrier Proteins / isolation & purification
                • Carrier Proteins / metabolism
                • Chromatography, Affinity
                • Chromatography, High Pressure Liquid
                • Gelatin / metabolism
                • Glycoproteins / chemistry
                • Glycoproteins / isolation & purification
                • Glycoproteins / metabolism
                • Glycosylation
                • Heparin / metabolism
                • Horses
                • Lectins
                • Male
                • Molecular Sequence Data
                • Oligosaccharides / chemistry
                • Oligosaccharides / isolation & purification
                • Prostatic Secretory Proteins
                • Proteins / chemistry
                • Semen / metabolism
                • Seminal Plasma Proteins
                • Sequence Homology, Amino Acid

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                Citations

                This article has been cited 8 times.
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                2. Sudheer Kumar C, Swamy MJ. Modulation of chaperone-like and membranolytic activities of major horse seminal plasma protein HSP-1/2 by L-carnitine. J Biosci 2017 Sep;42(3):469-479.
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                3. 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.
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                4. 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.
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