Metabolon disruption: a mechanism that regulates bicarbonate transport.
Carbonic anhydrases (CA) catalyze the reversible conversion of CO2 to HCO3-. Some bicarbonate transporters bind CA, forming a complex called a transport metabolon, to maximize the coupled catalytic/transport flux. SLC26A6, a plasma membrane Cl-/HCO3- exchanger with a suggested role in pancreatic HCO3- secretion, was found to bind the cytoplasmic enzyme CAII. Mutation of the identified CAII binding (CAB) site greatly reduced SLC26A6 activity, demonstrating the importance of the interaction. Regulation of SLC26A6 bicarbonate transport by protein kinase C (PKC) was investigated. Angiotensin II (AngII), which activates PKC, decreased Cl-/HCO3- exchange in cells coexpressing SLC26A6 and AT1a-AngII receptor. Activation of PKC reduced SLC26A6/CAII association in immunoprecipitates. Similarly, PKC activation displaced CAII from the plasma membrane, as monitored by immunofluorescence. Finally, mutation of a PKC site adjacent to the SLC26A6 CAB site rendered the transporter unresponsive to PKC. PKC therefore reduces CAII/SLC26A6 interaction, reducing bicarbonate transport rate. Taken together, our data support a mechanism for acute regulation of membrane transport: metabolon disruption.
Alvarez
Bernardo V
BV
Membrane Protein Research Group, Department of Physiology, University of Alberta, Edmonton, Alberta, Canada.
Vilas
Gonzalo L
GL
Casey
Joseph R
JR
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000235:genetics / Q000378:metabolism; D001639:Bicarbonates / Q000378:metabolism*; D001693:Biological Transport, Active / Q000235:genetics; D024402:Carbonic Anhydrase II / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D000076988:Sulfate Transporters; D013997:Time Factors; D014162:Transfection
D001639:Bicarbonates; D026901:Membrane Transport Proteins; C439529:SLC26A6 protein, human; D000076988:Sulfate Transporters; D000804:Angiotensin II; D011493:Protein Kinase C; D024402:Carbonic Anhydrase II