Characteristics of acid phosphatase from rainbow trout (Oncorhynchus mykiss) spermatozoa

• Autorzy: Sarosiek B. , Wysocka J. , Wysocki P. , Glogowski J.

Warianty tytułu

PL

Charakterystyka fosfatazy kwaśnej z plemników pstrąga tęczowego (Oncorhynchus mykiss)

• Języki publikacji: EN

Abstrakty

EN

Acid phosphatase (AcP) is a commonly observed enzyme in animal semen. In this study, AcP in rainbow trout (Oncorhynchus mykiss) spermatozoa was partly purified and characterized. Extraction in 0.85% NaCl with 0.1% Triton X-100 enabled obtaining 95% of total AcP activity observed in sperm supernatant. Kinetic characteristics were described for the enzyme from sperm extract and for the partly purified enzyme following gel filtration. The optimum pH was 5.8 for unpurified and 5.6 for partly purified enzyme. The affinity of the substrates measured in the sperm extract for p-nitrophenylphosphate dissodium salt and b-glycerophosphate was Km = 1.5 × 10-3M and Km = 1.9 × 10-3M, respectively. The Km for partly purified enzyme was similar at 1.67 × 10-3M measured with p-nitrophenylphosphate dissodium salt. L-tartaric acid and ammonium molybdate were the inhibitors of AcP for unpurified and partly purified enzyme. SDS-PAGE electrophoresis revealed that AcP from rainbow trout had a molecular weight of about 41 kDa.

PL

Kwaśna fosfataza (AcP) jest enzymem powszechnie występującym w nasieniu wielu gatunkóv zwierząt. W niniejszych badaniach nasienie pobrane od pstrąga tęczowego Oncorhynchus mykis odwirowa no, a następnie plemniki poddano ekstrakcji w 0,85% NaCl z 0,1% Tritonem X-100. Charakterystykę kine tyczną przeprowadzono dla nieoczyszczonego enzymu, a także dla częściowo oczyszczonego białka uzyskanego po filtracji żelowej. Dla enzymu z ekstraktu optimum pH wynosiło 5,8 z użyciem p-nitrofenylophosfo ranu jako substratu oraz 6,0 z użyciem β-glicerofosforanu (rys. 2). Dla enzymu po filtracji optimum ph wynosiło 5,6 (rys. 9). Powinowactwo do substratu określono przy użyciu p-nitrofenylophosforanu oraz β-glicerofosforanu, Km wynosiło odpowiednio: 1,5 x 10⁻³M i 1.9 x 10⁻³M dla enzymu z ekstraktu plemników Km dla częściowo oczyszczonej AcP wynosiło 1.67 x 10⁻³M, z użyciem p-nitrofenylophosforanu. Kwas winowy i molibdenian amonu okazały się inhibitorami badanej fosfatazy (rys. 5, 10). Elektroforeza SDS-PAGE pozwoliła na stwierdzenie, że enzym posiada masę cząsteczkową 41 kDa (rys. 7, 8).

Słowa kluczowe

EN

acid phosphatase; fish; rainbow trout; Oncorhynchus mykiss; spermatozoon; analytical procedure; sperm extract

• Wydawca: -
• Czasopismo: Archives of Polish Fisheries
• Rocznik: 2005
• Tom: 13
• Numer: 2
• Opis fizyczny: p.131-146,fig.,ref.

Twórcy

autor

Sarosiek B.

• Molecular Andrology Group, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Bydgoska 1/8, 10-243 Olsztyn, Poland

autor

Wysocka J.

autor

Wysocki P.

autor

Glogowski J.

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