Carbohydrate Affinity Chromatography Indicates That Arylsulfatase-a From Capacitated Boar Sperm Has Mannose and N-Acetylglucosamine/Sialic Acid Residues

Abstract

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.

Effects of Atrazine and Fenoxaprop-Ethyl on Capacitation and the Acrosomal Reaction in Boar Sperm

“…Capacitation and AR data shown in this study agree with results previously obtained in our lab and others. 39,40 PIAR values are similar to those reported by other authors. 28,31,37,[41][42][43] With 12 mM FE, the proportion of capacitated sperm diminished significantly compared with controls, whereas the noncapacitated proportion did not change.…”

Perfluorooctane Sulfonate (PFOS) and Perfluorohexane Sulfonate (PFHxS) Alters Protein Phosphorylation, Increase ROS Levels and DNA Fragmentation during In Vitro Capacitation of Boar Spermatozoa

“…The in vitro boar sperm capacitation methodology gave optimal results, obtaining levels of capacitation, similar to others previously reported [ 27 , 36 ]. The CTC technique is useful for differentiating among non-capacitated, capacitated, and AR spermatozoa in normal in vitro sperm, based on the CTC fluorescence patterns when they are in contact with intracellular Ca 2+ bound to membrane proteins [ 37 ].…”

Sialylation: fate decision of mammalian sperm development, fertilization, and male fertility

“…Sperm structure and sialylation on the membrane Sialylation is a widespread modification found on the cell surface [59] and is particularly abundant on the sperm plasma membrane [60][61][62][63]. Sperm can be divided into three regions: head, neck, and tail, each responsible for different functions [64].…”