Subcellular localization of adenylate cyclase of buffalo spermatozoa.
The intracellular localization of adenylate cyclase and 3',5'-cyclic nucleotide phosphodiesterase in buffalo sperm was examined. Adenylate cyclase activity is distributed in heads (8.4%), midpieces (16.6%), tails (49.5%) and 5.7% in the soluble supernatant; the total recovery being 81%. A 4-fold increase in specific activity was observed in the tail fraction relative to sonicated suspension. Further fractionation of the tail fraction into plasma membrane and microtubules by dialysis against low ionic strength buffer was followed by marker enzymes (Mg2+ -ATPase, 5'-nucleotidase and alkaline phosphatase) as well as by examination of fractions under electron microscope. The recovered adenylate cyclase (79%) was found in microtubules (45%) and plasma membrane (34%). Cyclic nucleotide phosphodiesterase in tails was distributed in tail plasma membrane (13.7%), microtubules (31.5%) and cytosol (34%) with a total recovery of 80%. Similar results were obtained when the distribution of adenylate cyclase and cyclic nucleotide phosphodiesterase was studied by treatment with Triton X-100; 40% activity of adenylate cyclase present in tails (about 20% relative to sperm sonicate) appeared in the soluble form by this method. The results are discussed in relation to control of cyclic AMP levels in buffalo sperm by adenylate cyclase and cyclic nucleotide phosphodiesterase.
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000032:analysis; D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases / Q000032:analysis; D000262:Adenylyl Cyclases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002020:Buffaloes; D017301:Ca(2+) Mg(2+)-ATPase; D002458:Cell Fractionation; D008297:Male; D008854:Microscopy, Electron; D009708:Nucleotidases / Q000032:analysis; D013094:Spermatozoa / Q000201:enzymology* / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000201:enzymology
D009708:Nucleotidases; D015720:5'-Nucleotidase; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D000262:Adenylyl Cyclases