Abstract
It was shown earlier that a 67-kDa protein purified from mouse kidney using polyclonal antibodies against melittin (a peptide from bee venom) interacted with Na,K-ATPase from rabbit kidney. In this study, a 43-kDa proteolytic fragment of Na,K-ATPase α-subunit interacting with the 67-kDa melittin-like protein was found. The α-subunit was hydrolyzed by trypsin in the presence of 0.5 mM ouabain (E2-conformation of Na,K-ATPase). A proteolytic fragment interacting with the 67-kDa melittin-like protein that was identified by mass-spectrometry is a region of the cytoplasmic domain of Na,K-ATPase α-subunit located between amino acid residues 591 and 775. The fragment includes a conservative DPPRA motif that occurs in many P-type ATPases. It was shown earlier that this motif of H,K-ATPase from gastric mucosa binds to melittin. We suggest that namely this motif of P-type ATPases is able to interact with proteins containing melittin-like modules.
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Abbreviations
- CHAPS:
-
3-[(3-cholamidoprolyl)dimethylammonium]-1-propanesulfonate
- ELISA:
-
enzyme-linked immunosorbent assay
- PMSF:
-
phenylmethylsulfonyl fluoride
- PBS:
-
phosphate-buffered saline
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Published in Russian in Biokhimiya, 2016, Vol. 81, No. 3, pp. 369–375.
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Kamanina, Y.V., Klimanova, E.A., Dergousova, E.A. et al. Identification of a region of the polypeptide chain of Na,K-ATPase α-subunit interacting with 67-kDa melittin-like protein. Biochemistry Moscow 81, 249–254 (2016). https://doi.org/10.1134/S000629791603007X
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DOI: https://doi.org/10.1134/S000629791603007X