Biochemical and conformational characterisation of HSP-3, a stallion seminal plasma protein of the cysteine-rich secretory protein (CRISP) family.
Abstract: HSP-3 is a member of the cysteine-rich secretory protein (CRISP) family from stallion seminal plasma. We report a large-scale purification protocol for native HSP-3. This protein is a non-glycosylated polypeptide chain with a pI of 8-9 and an isotope-averaged molecular mass of 24987 +/- 3 Da. The molecular mass of HSP-3, determined by equilibrium sedimentation, is 26 kDa, showing that the protein exists in solution as a monomer. The concentration of HSP-3 in the seminal plasma of different stallions ranged from 0.3 to 1.3 mg/ml. On average, 0.9-9 million HSP-3 molecules/cell coat the postacrosomal and mid-piece regions of an ejaculated, washed stallion spermatozoon, suggesting a role in sperm physiology. Conformational characterisation of purified HSP-3 was assessed by combination of circular dichroism and Fourier-transform infrared spectroscopies and differential scanning microcalorimetry. Based on secondary structure assignment, HSP-3 may belong to the alpha+beta class of proteins. Thermal denaturation of HSP-3 is irreversible and follows a non-two state transition characterised by a Tm of 64 degrees C, an enthalpy change of 75 kcal/mol, and a van 't Hoff enthalpy of 184 kcal/mol. Analysis of the spectroscopic and calorimetric data indicates the occurrence of aggregation of denatured HSP-3 molecules and suggests the monomer as the cooperative unfolding unit.
Publication Date: 1998-02-12 PubMed ID: 9459306DOI: 10.1016/s0014-5793(97)01514-7Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article explores a protein called HSP-3 found in stallion seminal plasma. This protein is a non-glycosylated polypeptide chain that could play a role in horse sperm physiology. The team developed a way to purify this protein and used various methods to study its characteristics and behaviour.
Significance of HSP-3
- HSP-3 is part of the CRISP family of proteins, which are found in the seminal plasma of stallions.
- The research examines the molecular structure and behavior of HSP-3, which could offer insights into its function in the semen of horses.
Purification Process
- The team developed a large-scale protocol for purifying HSP-3 in its native state.
- This purification process was critical for accurately analyzing the protein for further experimentation.
Characteristics of HSP-3
- The protein is a non-glycosylated polypeptide, meaning it does not contain sugar compounds attached.
- The isotope-averaged molecular mass of HSP-3 was determined to be 24987 +/- 3 Da.
- HSP-3 exists as a monomer or single molecule in solution as established by sedimentation testing.
- The concentration of HSP-3 in seminal plasma varied among stallions. The range was found to be from 0.3 to 1.3 mg/ml.
Role in Sperm Physiology
- The researchers noted that an average of 0.9 to 9 million HSP-3 molecules per cell coat specific regions of a washed, ejaculated stallion sperm, indicating a potential role in sperm physiology.
Conformational Characterisation of HSP-3
- The conformation (shape) and features of HSP-3 were investigated using circular dichroism and Fourier-transform infrared spectroscopies, and differential scanning microcalorimetry.
- HSP-3 is likely part of the alpha+beta class of proteins based on secondary structure assignment.
- Thermal denaturation of HSP-3 follows a non-two state transition, meaning it undergoes complex changes in structure and properties when heated.
- Spectroscopic and calorimetric data suggest that there is aggregation of HSP-3 molecules after denaturation, with the monomer acting as the cooperative unfolding unit.
Cite This Article
APA
Magdaleno L, Gasset M, Varea J, Schambony AM, Urbanke C, Raida M, Töpfer-Petersen E, Calvete JJ.
(1998).
Biochemical and conformational characterisation of HSP-3, a stallion seminal plasma protein of the cysteine-rich secretory protein (CRISP) family.
FEBS Lett, 420(2-3), 179-185.
https://doi.org/10.1016/s0014-5793(97)01514-7 Publication
Researcher Affiliations
- Instituto de Química-Física Rocasolano, C.S.I.C., Madrid, Spain.
MeSH Terms
- Animals
- Calorimetry, Differential Scanning
- Chromatography, Gel
- Circular Dichroism
- Electrophoresis, Polyacrylamide Gel
- Fluorescent Antibody Technique
- Horses
- Isoelectric Point
- Male
- Mass Spectrometry
- Molecular Weight
- Prostatic Secretory Proteins
- Protein Conformation
- Protein Denaturation
- Protein Folding
- Protein Structure, Secondary
- Proteins / chemistry
- Proteins / isolation & purification
- Seminal Plasma Proteins
- Spectroscopy, Fourier Transform Infrared
- Spermatozoa / chemistry
- Spermatozoa / cytology
- Temperature
- Thermodynamics
Citations
This article has been cited 6 times.- Gonzalez SN, Sulzyk V, Weigel Muñoz M, Cuasnicu PS. Cysteine-Rich Secretory Proteins (CRISP) are Key Players in Mammalian Fertilization and Fertility.. Front Cell Dev Biol 2021;9:800351.
- Bubenickova F, Postlerova P, Simonik O, Sirohi J, Sichtar J. Effect of Seminal Plasma Protein Fractions on Stallion Sperm Cryopreservation.. Int J Mol Sci 2020 Sep 3;21(17).
- Tadokoro T, Modahl CM, Maenaka K, Aoki-Shioi N. Cysteine-Rich Secretory Proteins (CRISPs) From Venomous Snakes: An Overview of the Functional Diversity in A Large and Underappreciated Superfamily.. Toxins (Basel) 2020 Mar 12;12(3).
- Johannisson A, Al-Essawe EM, Al-Saffar AK, Karkehabadi S, Lima-Verde I, Wulf M, Aurich C, Morrell JM. Season does not have a deleterious effect on proportions of stallion seminal plasma proteins.. J Reprod Dev 2020 Jun 12;66(3):215-221.
- Wilbers RHP, Schneiter R, Holterman MHM, Drurey C, Smant G, Asojo OA, Maizels RM, Lozano-Torres JL. Secreted venom allergen-like proteins of helminths: Conserved modulators of host responses in animals and plants.. PLoS Pathog 2018 Oct;14(10):e1007300.
- Koppers AJ, Reddy T, O'Bryan MK. The role of cysteine-rich secretory proteins in male fertility.. Asian J Androl 2011 Jan;13(1):111-7.
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