Assembly of the extracellular domain of the Na,K-ATPase beta subunit with the alpha subunit. Analysis of beta subunit chimeras and carboxyl-terminal deletions.
The role of the extracellular domain of the Na,K-ATPase beta subunit in assembly with the alpha subunit was investigated. A chimeric protein consisting of the extracellular domain of the beta subunit fused with the transmembrane and cytoplasmic domains of dipeptidyl peptidase IV assembles with the alpha subunit. An inverse chimera consisting of the cytoplasmic and transmembrane domains of the beta subunit fused with the extracellular domain of dipeptidyl peptidase IV does not assemble with the alpha subunit. The assembly data from these chimeras demonstrate that the extracellular domain of the beta subunit is both necessary and sufficient for assembly with the alpha subunit. Deletions of up to 146 extracellular amino acids from the carboxyl terminus of the beta subunit appear to result in misfolding of the subunit, but do allow reduced assembly with the alpha subunit. Together, the assembly data from chimeras and carboxyl-terminal deletions have identified a 96-residue extracellular domain which contains sequences involved in subunit assembly. While the chimeric subunits properly localize to the plasma membrane, deletion of as few as 4 amino acids from the carboxyl terminus impairs the ability of the beta subunit to be transported to the plasma membrane.
Hamrick
M
M
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Renaud
K J
KJ
Fambrough
D M
DM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D018076:DNA, Complementary; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4; D000254:Sodium-Potassium-Exchanging ATPase