TOPOLOGY OF NA,K-ATPASE ALPHA-SUBUNIT EPITOPES ANALYZED WITH OLIGOPEPTIDE-SPECIFIC ANTIBODIES AND DOUBLE-LABELING IMMUNOELECTRON MICROSCOPY

被引：23

作者：

NING, G

MAUNSBACH, AB

LEE, YJ

MOLLER, JV

机构：

[1] UNIV AARHUS, INST BIOPHYS, DK-8000 AARHUS C, DENMARK

[2] UNIV AARHUS, BIOMEMBRANE RES CTR, DK-8000 AARHUS C, DENMARK

来源：

FEBS LETTERS | 1993年 / 336卷 / 03期

关键词：

NA; K-ATPASE; ALPHA-SUBUNIT; TOPOLOGY; MEMBRANE EPITOPE; DOUBLE LABELING; OLIGOPEPTIDE-SPECIFIC ANTIBODY; IMMUNOELECTRON MICROSCOPY;

D O I：

10.1016/0014-5793(93)80868-U

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Using four oligopeptide-specific polyclonal antibodies, we mapped the alpha subunit of Na,K-ATPase by double labeling immunoelectron microscopy combined with negative staining. The results show that the epitopes of the N-terminus (Gly(1)-His(13)), C-terminus Ile(1002)-Tyr(1016) and Leu(815)-Gln(828) are located on the same face of crystallized Na,K-ATPase membranes from pig kidney, whereas the epitope Asn(889)-Gln(903) is present on the opposite side. The present study demonstrates the cytoplasmic location of C-terminus and that Leu(815)-Gln(828) is exposed on the cytoplasmic and Asn(889)-Gln(903) on the extracellular side. The results are consistent with an eight- or ten-segment model, and support the existence of an M5/M6 loop and the presence of one transmembrane segment between Leu(815)-Gln(828) and Asn(889)-Gln(903).

引用

页码：521 / 524

页数：4

共 21 条

[1] EPITOPE MAPPING BY AMINO-ACID-SEQUENCE-SPECIFIC ANTIBODIES REVEALS THAT BOTH ENDS OF THE ALPHA-SUBUNIT OF NA+/K+-ATPASE ARE LOCATED ON THE CYTOPLASMIC SIDE OF THE MEMBRANE [J].