Characterization of β-N-acetylglucosaminidase from human epididymis
β-N-acetylglucosaminidase (NAG) activity in human epididymal fluid was separated into two forms (I and II) after HPLC-hydrophobic interaction chromatography. Both forms exhibited maximal activity at a pH of around 4.5 and had a molecular weight of 125 kD when determined by Superose-HPLC. After incub...
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1995
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01056263_v18_n5_p263_Miranda http://hdl.handle.net/20.500.12110/paper_01056263_v18_n5_p263_Miranda |
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paper:paper_01056263_v18_n5_p263_Miranda2023-06-08T15:10:33Z Characterization of β-N-acetylglucosaminidase from human epididymis β-N-acetylglucosaminidase Epididymis Glycosidase Hexosaminidase Human Sperm maturation beta n acetylhexosaminidase adult aged article cell membrane electrophoresis electrophoretic mobility enzyme activity enzyme localization epididymis high performance liquid chromatography human human cell human tissue male molecular weight ph priority journal seminal plasma spermatozoon Acetylglucosaminidase Aged Aged, 80 and over Chromatography, Gel Chromatography, High Pressure Liquid Cytosol Epididymis Human Hydrogen-Ion Concentration Isoenzymes Kinetics Male Middle Age Molecular Weight Orchiectomy Prostatic Neoplasms Spermatozoa Subcellular Fractions Support, Non-U.S. Gov't β-N-acetylglucosaminidase (NAG) activity in human epididymal fluid was separated into two forms (I and II) after HPLC-hydrophobic interaction chromatography. Both forms exhibited maximal activity at a pH of around 4.5 and had a molecular weight of 125 kD when determined by Superose-HPLC. After incubation at 50°C, form I retained only 30% of its activity while form II retained 90% activity. When analysed by non-denaturing electrophoresis, form I displayed higher electrophoretic mobility than did form II. These features indicate that the I and II isoforms found in the human epididymis are the A and B forms present in other tissues. NAG activity was measured in the fluid obtained form the different epididymal regions of 13 different samples. An average four-fold increase in activity between the proximal caput and distal corpus was found. The contribution of each isoform to the total activity was studied. The proximal caput found to be rich in the A isoform (59%), whereas the B form was predominant in the distal corpus (65%). Human spermatozoa contain membrane-associated NAG activity with an isoform distribution similar to that found in cauda epididymal fluid (CEP, 80% B). Finally, enzyme activity in CEP was two-fold greater than in seminal plasma. Taken together these results suggest that NAG may become associated with human spermatozoa during epididymal transit. 1995 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01056263_v18_n5_p263_Miranda http://hdl.handle.net/20.500.12110/paper_01056263_v18_n5_p263_Miranda |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
β-N-acetylglucosaminidase Epididymis Glycosidase Hexosaminidase Human Sperm maturation beta n acetylhexosaminidase adult aged article cell membrane electrophoresis electrophoretic mobility enzyme activity enzyme localization epididymis high performance liquid chromatography human human cell human tissue male molecular weight ph priority journal seminal plasma spermatozoon Acetylglucosaminidase Aged Aged, 80 and over Chromatography, Gel Chromatography, High Pressure Liquid Cytosol Epididymis Human Hydrogen-Ion Concentration Isoenzymes Kinetics Male Middle Age Molecular Weight Orchiectomy Prostatic Neoplasms Spermatozoa Subcellular Fractions Support, Non-U.S. Gov't |
| spellingShingle |
β-N-acetylglucosaminidase Epididymis Glycosidase Hexosaminidase Human Sperm maturation beta n acetylhexosaminidase adult aged article cell membrane electrophoresis electrophoretic mobility enzyme activity enzyme localization epididymis high performance liquid chromatography human human cell human tissue male molecular weight ph priority journal seminal plasma spermatozoon Acetylglucosaminidase Aged Aged, 80 and over Chromatography, Gel Chromatography, High Pressure Liquid Cytosol Epididymis Human Hydrogen-Ion Concentration Isoenzymes Kinetics Male Middle Age Molecular Weight Orchiectomy Prostatic Neoplasms Spermatozoa Subcellular Fractions Support, Non-U.S. Gov't Characterization of β-N-acetylglucosaminidase from human epididymis |
| topic_facet |
β-N-acetylglucosaminidase Epididymis Glycosidase Hexosaminidase Human Sperm maturation beta n acetylhexosaminidase adult aged article cell membrane electrophoresis electrophoretic mobility enzyme activity enzyme localization epididymis high performance liquid chromatography human human cell human tissue male molecular weight ph priority journal seminal plasma spermatozoon Acetylglucosaminidase Aged Aged, 80 and over Chromatography, Gel Chromatography, High Pressure Liquid Cytosol Epididymis Human Hydrogen-Ion Concentration Isoenzymes Kinetics Male Middle Age Molecular Weight Orchiectomy Prostatic Neoplasms Spermatozoa Subcellular Fractions Support, Non-U.S. Gov't |
| description |
β-N-acetylglucosaminidase (NAG) activity in human epididymal fluid was separated into two forms (I and II) after HPLC-hydrophobic interaction chromatography. Both forms exhibited maximal activity at a pH of around 4.5 and had a molecular weight of 125 kD when determined by Superose-HPLC. After incubation at 50°C, form I retained only 30% of its activity while form II retained 90% activity. When analysed by non-denaturing electrophoresis, form I displayed higher electrophoretic mobility than did form II. These features indicate that the I and II isoforms found in the human epididymis are the A and B forms present in other tissues. NAG activity was measured in the fluid obtained form the different epididymal regions of 13 different samples. An average four-fold increase in activity between the proximal caput and distal corpus was found. The contribution of each isoform to the total activity was studied. The proximal caput found to be rich in the A isoform (59%), whereas the B form was predominant in the distal corpus (65%). Human spermatozoa contain membrane-associated NAG activity with an isoform distribution similar to that found in cauda epididymal fluid (CEP, 80% B). Finally, enzyme activity in CEP was two-fold greater than in seminal plasma. Taken together these results suggest that NAG may become associated with human spermatozoa during epididymal transit. |
| title |
Characterization of β-N-acetylglucosaminidase from human epididymis |
| title_short |
Characterization of β-N-acetylglucosaminidase from human epididymis |
| title_full |
Characterization of β-N-acetylglucosaminidase from human epididymis |
| title_fullStr |
Characterization of β-N-acetylglucosaminidase from human epididymis |
| title_full_unstemmed |
Characterization of β-N-acetylglucosaminidase from human epididymis |
| title_sort |
characterization of β-n-acetylglucosaminidase from human epididymis |
| publishDate |
1995 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01056263_v18_n5_p263_Miranda http://hdl.handle.net/20.500.12110/paper_01056263_v18_n5_p263_Miranda |
| _version_ |
1768542309006180352 |