FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Biochem J / Amino acid sequence of HSP-1, a major protein of stallion se...
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.
Get Full Text
Publication Date: 1995-09-01 PubMed ID: 7654203PubMed Central: PMC1135940DOI: 10.1042/bj3100615Google 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.
LinkedInCopy
  • 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

                References

                This article includes 25 references
                1. Calvete JJ, Raida M, Sanz L, Wempe F, Scheit KH, Romero A, Töpfer-Petersen E. Localization and structural characterization of an oligosaccharide O-linked to bovine PDC-109. Quantitation of the glycoprotein in seminal plasma and on the surface of ejaculated and capacitated spermatozoa.. FEBS Lett 1994 Aug 22;350(2-3):203-6.
                  pubmed: 8070564doi: 10.1016/0014-5793(94)00768-3google scholar: lookup
                2. Haselbeck A, Schickaneder E, von der Eltz H, Hösel W. Structural characterization of glycoprotein carbohydrate chains by using diagoxigenin-labeled lectins on blots.. Anal Biochem 1990 Nov 15;191(1):25-30.
                  pubmed: 2127661doi: 10.1016/0003-2697(90)90381-igoogle scholar: lookup
                3. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 1970 Aug 15;227(5259):680-5.
                  pubmed: 5432063doi: 10.1038/227680a0google scholar: lookup
                4. Sanz L, Calvete JJ, Mann K, Gabius HJ, Töpfer-Petersen E. Isolation and biochemical characterization of heparin-binding proteins from boar seminal plasma: a dual role for spermadhesins in fertilization.. Mol Reprod Dev 1993 May;35(1):37-43.
                  pubmed: 8507478doi: 10.1002/mrd.1080350107google scholar: lookup
                5. Miller DJ, Winer MA, Ax RL. Heparin-binding proteins from seminal plasma bind to bovine spermatozoa and modulate capacitation by heparin.. Biol Reprod 1990 May-Jun;42(5-6):899-915.
                  pubmed: 2383614doi: 10.1095/biolreprod42.6.899google scholar: lookup
                6. Skorstengaard K, Jensen MS, Sahl P, Petersen TE, Magnusson S. Complete primary structure of bovine plasma fibronectin.. Eur J Biochem 1986 Dec 1;161(2):441-53.
                  pubmed: 3780752doi: 10.1111/j.1432-1033.1986.tb10464.xgoogle scholar: lookup
                7. Anumula KR, Taylor PB. Rapid characterization of asparagine-linked oligosaccharides isolated from glycoproteins using a carbohydrate analyzer.. Eur J Biochem 1991 Jan 1;195(1):269-80.
                  pubmed: 1991474doi: 10.1111/j.1432-1033.1991.tb15703.xgoogle scholar: lookup
                8. Collier IE, Krasnov PA, Strongin AY, Birkedal-Hansen H, Goldberg GI. Alanine scanning mutagenesis and functional analysis of the fibronectin-like collagen-binding domain from human 92-kDa type IV collagenase.. J Biol Chem 1992 Apr 5;267(10):6776-81.
                  pubmed: 1313021
                9. Bányai L, Trexler M, Koncz S, Gyenes M, Sipos G, Patthy L. The collagen-binding site of type-II units of bovine seminal fluid protein PDC-109 and fibronectin.. Eur J Biochem 1990 Nov 13;193(3):801-6.
                  pubmed: 2249694doi: 10.1111/j.1432-1033.1990.tb19403.xgoogle scholar: lookup
                10. Aumüller G, Vesper M, Seitz J, Kemme M, Scheit KH. Binding of a major secretory protein from bull seminal vesicles to bovine spermatozoa.. Cell Tissue Res 1988 May;252(2):377-84.
                  pubmed: 3383217doi: 10.1007/BF00214380google scholar: lookup
                11. Chandonnet L, Roberts KD, Chapdelaine A, Manjunath P. Identification of heparin-binding proteins in bovine seminal plasma.. Mol Reprod Dev 1990 Aug;26(4):313-8.
                  pubmed: 2171588doi: 10.1002/mrd.1080260404google scholar: lookup
                12. Leblond E, Desnoyers L, Manjunath P. Phosphorylcholine-binding proteins from the seminal fluids of different species share antigenic determinants with the major proteins of bovine seminal plasma.. Mol Reprod Dev 1993 Apr;34(4):443-9.
                  pubmed: 7682422doi: 10.1002/mrd.1080340414google scholar: lookup
                13. Varner DD, Bowen JA, Johnson L. Effect of heparin on capacitation/acrosome reaction of equine sperm.. Arch Androl 1993 Nov-Dec;31(3):199-207.
                  pubmed: 8274046doi: 10.3109/01485019308988400google scholar: lookup
                14. Wieland F, Heitzer R, Schaefer W. Asparaginylglucose: Novel type of carbohydrate linkage.. Proc Natl Acad Sci U S A 1983 Sep;80(18):5470-4.
                  pubmed: 16593364doi: 10.1073/pnas.80.18.5470google scholar: lookup
                15. Einspanier R, Krause I, Calvete JJ, Töfper-Petersen E, Klostermeyer H, Karg H. Bovine seminal plasma aSFP: localization of disulfide bridges and detection of three different isoelectric forms.. FEBS Lett 1994 May 9;344(1):61-4.
                  pubmed: 8181566doi: 10.1016/0014-5793(94)00362-9google scholar: lookup
                16. Esch FS, Ling NC, Böhlen P, Ying SY, Guillemin R. Primary structure of PDC-109, a major protein constituent of bovine seminal plasma.. Biochem Biophys Res Commun 1983 Jun 29;113(3):861-7.
                  pubmed: 6870895doi: 10.1016/0006-291x(83)91078-1google scholar: lookup
                17. Jonákova V, Sanz L, Calvete JJ, Henschen A, Cechová D, Töpfer-Petersen E. Isolation and biochemical characterization of a zona pellucida-binding glycoprotein of boar spermatozoa.. FEBS Lett 1991 Mar 11;280(1):183-6.
                  pubmed: 1849093doi: 10.1016/0014-5793(91)80233-sgoogle scholar: lookup
                18. Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX.. Nucleic Acids Res 1984 Jan 11;12(1 Pt 1):387-95.
                  pubmed: 6546423doi: 10.1093/nar/12.1part1.387google scholar: lookup
                19. Manjunath P, Chandonnet L, Leblond E, Desnoyers L. Major proteins of bovine seminal vesicles bind to spermatozoa.. Biol Reprod 1994 Jan;50(1):27-37.
                  pubmed: 8312447doi: 10.1095/biolreprod50.1.27google scholar: lookup
                20. Manjunath P, Sairam MR. Purification and biochemical characterization of three major acidic proteins (BSP-A1, BSP-A2 and BSP-A3) from bovine seminal plasma.. Biochem J 1987 Feb 1;241(3):685-92.
                  pubmed: 3593217doi: 10.1042/bj2410685google scholar: lookup
                21. Desnoyers L, Manjunath P. Major proteins of bovine seminal plasma exhibit novel interactions with phospholipid.. J Biol Chem 1992 May 15;267(14):10149-55.
                  pubmed: 1577785
                22. Parrish JJ, Susko-Parrish JL, Handrow RR, Ax RL, First NL. Effect of sulfated glycoconjugates on capacitation and the acrosome reaction of bovine and hamster spermatozoa.. Gamete Res 1989 Dec;24(4):403-13.
                  pubmed: 2480320doi: 10.1002/mrd.1120240407google scholar: lookup
                23. Seidah NG, Manjunath P, Rochemont J, Sairam MR, Chrétien M. Complete amino acid sequence of BSP-A3 from bovine seminal plasma. Homology to PDC-109 and to the collagen-binding domain of fibronectin.. Biochem J 1987 Apr 1;243(1):195-203.
                  pubmed: 3606570doi: 10.1042/bj2430195google scholar: lookup
                24. Valencia A, Wens MA, Merchant H, Reyes R, Delgado NM. Capacitation of human spermatozoa by heparin.. Arch Androl 1984;12 Suppl:109-13.
                  pubmed: 6535449
                25. Sanz L, Calvete JJ, Mann K, Schäfer W, Schmid ER, Amselgruber W, Sinowatz F, Ehrhard M, Töpfer-Petersen E. The complete primary structure of the spermadhesin AWN, a zona pellucida-binding protein isolated from boar spermatozoa.. FEBS Lett 1992 Apr 6;300(3):213-8.
                  pubmed: 1555646doi: 10.1016/0014-5793(92)80848-bgoogle scholar: lookup

                Citations

                This article has been cited 8 times.
                1. Defaus S, Avilés M, Andreu D, Gutiérrez-Gallego R. Lectin-Binding Specificity of the Fertilization-Relevant Protein PDC-109 by Means of Surface Plasmon Resonance and Carbohydrate REcognition Domain EXcision-Mass Spectrometry.. Int J Mol Sci 2018 Apr 4;19(4).
                  doi: 10.3390/ijms19041076pubmed: 29617298google scholar: lookup
                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.
                  doi: 10.1007/s12038-017-9693-6pubmed: 29358560google scholar: lookup
                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.
                  doi: 10.1186/s12958-015-0093-1pubmed: 26272219google scholar: lookup
                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.
                  doi: 10.1186/s12958-015-0058-4pubmed: 26084664google scholar: lookup
                5. Anbazhagan V, Sankhala RS, Singh BP, Swamy MJ. Isothermal titration calorimetric studies on the interaction of the major bovine seminal plasma protein, PDC-109 with phospholipid membranes.. PLoS One 2011;6(10):e25993.
                  doi: 10.1371/journal.pone.0025993pubmed: 22022488google scholar: lookup
                6. Manjunath P, Lefebvre J, Jois PS, Fan J, Wright MW. New nomenclature for mammalian BSP genes.. Biol Reprod 2009 Mar;80(3):394-7.
                  doi: 10.1095/biolreprod.108.074088pubmed: 18923155google scholar: lookup
                7. Villemure M, Lazure C, Manjunath P. Isolation and characterization of gelatin-binding proteins from goat seminal plasma.. Reprod Biol Endocrinol 2003 Apr 28;1:39.
                  doi: 10.1186/1477-7827-1-39pubmed: 12737634google scholar: lookup
                8. Bezouska K, Sklenár J, Novák P, Halada P, Havlícek V, Kraus M, Tichá M, Jonáková V. Determination of the complete covalent structure of the major glycoform of DQH sperm surface protein, a novel trypsin-resistant boar seminal plasma O-glycoprotein related to pB1 protein.. Protein Sci 1999 Jul;8(7):1551-6.
                  doi: 10.1110/ps.8.7.1551pubmed: 10422846google scholar: lookup
                Subscribe to our newsletter
                Join 99,359 horse owners like you who receive our email updates on horse health and nutrition.