Effect of Colchicine on Sensitivity of Duck Salt Gland Na,K-ATPase to Na⁺

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S. S. Yakushev, E. M. Kumskova, A. M. Rubtsov, and O. D. Lopina^*

Department of Biochemistry, Faculty of Biology, Lomonosov Moscow State University, 119991 Moscow, Russia; fax: (495) 939-3955; E-mail: od_lopina@mail.ru

^* To whom correspondence should be addressed.

Received November 26, 2007; Revision received February 27, 2008

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Low molecular mass proteins of the FXYD family that affect the sensitivity of Na,K-ATPase to Na⁺ and K⁺ are known to be present in Na,K-ATPases in various tissues. In particular, in Na,K-ATPase from kidney a gamma-subunit (with electrophoretic mobility corresponding to molecular mass of about 10 kD) is present, and Na,K-ATPase preparations from heart contain phospholemman (electrophoretic mobility of this protein corresponds to molecular mass of 13-14 kD), which provides for the interaction of heart Na,K-ATPase with cytoskeletal microtubules. Disruption of microtubules by colchicine removes phospholemman from heart Na,K-ATPase preparations. The goal of the present study was to reveal a low molecular mass protein (probably a member of FXYD family) in preparation of Na,K-ATPase from duck salt glands. Immunoprecipitation of solubilized duck salt gland Na,K-ATPase using antibodies against alpha1-subunit results in the coprecipitation of a 13 kD protein with the Na,K-ATPase complex. Treatment of homogenate from duck salt glands with colchicine removes this protein from the purified preparation of Na,K-ATPase. Simultaneously, we observed a decrease in the sensitivity of Na,K-ATPase to Na⁺ at pH 6.5. However, colchicine treatment of homogenate from rabbit kidney does not affect either the sensitivity of Na,K-ATPase obtained from this homogenate to Na⁺ or the content of 10 kD protein (presumably gamma-subunit). The data suggest that phospholemman (or a similar member of the FXYD family) tightly interacts with Na,K-ATPase from duck salt glands and binds it to microtubules, simultaneously participating in the regulation of the sensitivity of Na,K-ATPase to Na⁺.

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KEY WORDS: Na,K-ATPase, colchicine, FXYD family proteins, duck salt glands

DOI: 10.1134/S0006297908090058

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