Identification of a Region of the Polypeptide Chain of Na,K-ATPase α-Subunit Interacting with 67-kDa Melittin-Like Protein.
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.
Lomonosov Moscow State University, Department of Biochemistry, School of Biology, Moscow, 119991, Russia. od_lopina@mail.ru.
Yu V
YV
Kamanina
E A
EA
Klimanova
E A
EA
Dergousova
I Yu
IY
Petrushanko
O D
OD
Lopina
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D047468:Immunoprecipitation; D008555:Melitten / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000378:metabolism
D010455:Peptides; D021122:Protein Subunits; D008555:Melitten; D014357:Trypsin; D000254:Sodium-Potassium-Exchanging ATPase