Carbohydrate affinity chromatography indicates that arylsulfatase-A from capacitated boar sperm has mannose and N-acetylglucosamine/sialic acid residues.
Carbohydrate residues on membrane proteins from sperm are important in gamete interaction. In recent years, Arylsulfatase A (AS-A) has been acquiring an important role from the various putative gamete interaction responsibles in sperm. The aim of this study was to determine if the capacitated boar sperm Arylsulfatase-A (AS-A), contains D-mannose, N-acetylglucosamine and/or sialic acid residues by its purification using affinity chromatography with Concanavalia ensiformis Agglutinin(Con-A) or Wheat Germ Agglutinin (WGA) as ligands. Sperm samples were capacitated in TALP-HEPES medium. Protein extract was added to the affinity columns. Sequencing of retained proteins was done after SDS-PAGE. Total capacitated sperm proteins electrophoresis showed molecular masses between 14 kDa and 102 kDa. A major band of 68 kDa, and 2 minor bands of 52 kDa and 47 kDa were observed. They were AS-A, hyaluronidase and lactadherin, respectively. The Con-A-retained proteins (RP) pattern showed bands from 14 to 98 kDa. After sequencing and BLAST analysis, the 62 kDa band corresponded to Arylsulfatase-A. The WGA RP fraction showed bands from 14 to 100 kDa. The 65 kDa band corresponded to AS-A. This study showed that AS-A has mannose, N-acetylglucosamine and/or sialic acid residues as part of its glycosilation. In this study AS-A was isolated from boar capacitated sperm by affinity chromatography using separately Con-A and WGA, indicating that there are mannose, N-acetylglucosamine and/or sialic acid residues in its glycosilation. AS-A is a membrane protein of capacitated sperm. Further investigation is needed to fully characterize the glycosidic residues bore by AS-A and to determine its function.
Jiménez
Irma
I
Doctorado en Ciencias Biológicas, Universidad Autónoma Metropolitana, México.
Fierro
Reyna
R
González-Márquez
Humberto
H
Mendoza-Hernández
Guillermo
G
Romo
Salvador
S
Betancourt
Miguel
M
D000117:Acetylglucosamine / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D002553:Cerebroside-Sulfatase / Q000737:chemistry* / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D006595:Hexosamines / Q000032:analysis; D008297:Male; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D013075:Sperm Capacitation / Q000502:physiology*; D013094:Spermatozoa / Q000201:enzymology*; D013552:Swine
D006595:Hexosamines; C008322:mannosamine; D002553:Cerebroside-Sulfatase; D019158:N-Acetylneuraminic Acid; D000117:Acetylglucosamine