Identification and partial characterization of alpha-1,4-glucosidase activity in equine epididymal fluid.
The expression of alpha-1,4-glucosidase activity was fluorometrically and electrophoretically assessed in the epididymal fluid and seminal plasma of stallions. alpha-Glucosidase specific activity in the epididymis increased significantly from the proximal caput to the cauda. Stallion epididymal glucosidase maintained activity in a wide range of pH, with two distinct peaks (around pH 4.0 and 6.0, respectively). Enzyme activities at different pH, inhibition assays with sodium dodecyl sulfate (SDS) and maltotriose (MTT, selective inhibitors of alpha-glucosidases "acidic" and "neutral" isoforms, described in other tissues) and the electrophoretic analysis in native and native/SDS-PAGE conditions, indicated that stallion epididymal glucosidase was due to two catalytically active forms. These forms, analyzed by non-denaturing electrophoresis, exhibited different electrophoretic mobility and molecular weight. Samples from the proximal caput of the epididymis were rich in Form II or "neutral" form, whereas the "acid" or Form I seemed to be predominate in the cauda epididymal region. At physiological pH, Form II was predominant in the seminal plasma. The physiological role(s) of these forms is uncertain, but based on their ability to hydrolyze glucosidic linkage, they probably are involved in degradation/modifications of epididymal fluid and/or spermatozoa glycoconjugates, thereby participating in plasma membrane remodeling associated with sperm maturation.
Dias
Angelo J B
AJ
Setor Biologia da Reprodução, Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Av. Alberto Lamego 2000, Horto, Campos dos Goytacazes, RJ, Brazil.
Maia
Marcos S
MS
Retamal
Claudio A
CA
López
María Luisa
ML
D000818:Animals; D001826:Body Fluids / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004822:Epididymis / Q000201:enzymology*; D006736:Horses / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D012661:Semen / Q000201:enzymology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D014312:Trisaccharides / Q000494:pharmacology; D000520:alpha-Glucosidases / Q000032:analysis* / Q000378:metabolism
D004791:Enzyme Inhibitors; D014312:Trisaccharides; D012967:Sodium Dodecyl Sulfate; C008317:maltotriose; D000520:alpha-Glucosidases