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PMID:15647390

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Citation

Codina, J, Li, J and Dubose, TD Jr (2005) CD63 interacts with the carboxy terminus of the colonic H+-K+-ATPase to decrease [corrected] plasma membrane localization and 86Rb+ uptake. Am. J. Physiol., Cell Physiol. 288:C1279-86

Abstract

The carboxy terminus (CT) of the colonic H(+)-K(+)-ATPase is required for stable assembly with the beta-subunit, translocation to the plasma membrane, and efficient function of the transporter. To identify protein-protein interactions involved in the localization and function of HKalpha(2), we selected 84 amino acids in the CT of the alpha-subunit of mouse colonic H(+)-K(+)-ATPase (CT-HKalpha(2)) as the bait in a yeast two-hybrid screen of a mouse kidney cDNA library. The longest identified clone was CD63. To characterize the interaction of CT-HKalpha(2) with CD63, recombinant CT-HKalpha(2) and CD63 were synthesized in vitro and incubated, and complexes were immunoprecipitated. CT-HKalpha(2) protein (but not CT-HKalpha(1)) coprecipitated with CD63, confirming stable assembly of HKalpha(2) with CD63. In HEK-293 transfected with HKalpha(2) plus beta(1)-Na(+)-K(+)-ATPase, suppression of CD63 by RNA interference increased cell surface expression of HKalpha(2)/NKbeta(1) and (86)Rb(+) uptake. These studies demonstrate that CD63 participates in the regulation of the abundance of the HKalpha(2)-NKbeta(1) complex in the cell membrane.

Links

PubMed PMC1868892 Online version:10.1152/ajpcell.00463.2004

Keywords

Animals; Antigens, CD/metabolism; Antigens, CD/physiology; Antigens, CD63; Cell Line; Cell Membrane/enzymology; Colon/enzymology; Gene Expression/physiology; H(+)-K(+)-Exchanging ATPase/metabolism; Humans; Kidney/enzymology; Mice; Platelet Membrane Glycoproteins/metabolism; Platelet Membrane Glycoproteins/physiology; Rats; Rubidium Radioisotopes/diagnostic use; Saccharomyces cerevisiae

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:CD63

located_in

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CD63

acts_upstream_of_or_within

GO:0034613: cellular protein localization

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CD63

acts_upstream_of_or_within

GO:1901379: regulation of potassium ion transmembrane transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:CD63

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:Q9Z1W8

F

Seeded From UniProt

complete

MOUSE:CD63

acts_upstream_of_or_within

GO:0034613: cellular protein localization

ECO:0000266: sequence orthology evidence used in manual assertion

UniProtKB:P08962

P

Seeded From UniProt

complete

MOUSE:CD63

acts_upstream_of_or_within

GO:1901379: regulation of potassium ion transmembrane transport

ECO:0000266: sequence orthology evidence used in manual assertion

UniProtKB:P08962

P

Seeded From UniProt

complete

MOUSE:AT12A

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P41731

F

Seeded From UniProt

complete

HUMAN:CD63

part_of

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CD63

involved_in

GO:0034613: cellular protein localization

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CD63

involved_in

GO:1901379: regulation of potassium ion transmembrane transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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