Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Mutational analysis of histidine residues in the rabbit Na+/dicarboxylate co-transporter NaDC-1.
Pajor AM
,
Sun N
,
Valmonte HG
.
???displayArticle.abstract???
Succinate transport by the rabbit Na+/dicarboxylate co-transporter, NaDC-1, expressed in Xenopus oocytes was inhibited by the histidyl-selective reagent diethyl pyrocarbonate (DEPC). Therefore the role of histidine residues in the function of NaDC-1 was examined by site-directed mutagenesis. All 11 histidine residues in NaDC-1 were converted to alanine, but only mutant H106A exhibited a decrease in succinate transport. Additional mutations of NaDC-1 at position 106 showed that aspartic acid and asparagine, but not arginine, can substitute for histidine. Examination of succinate and citrate kinetics of H106A revealed a decrease in Vmax with no change in Km. Cell surface biotinylation experiments showed that the transport activity of all four mutants at position 106 was correlated with the amount of cell surface expression, suggesting a role of His-106 in membrane expression rather than function. Two of the histidine mutants, H153A and H569A, exhibited insensitivity to inhibition by DEPC, indicating that these residues are involved in binding DEPC. Neither of these residues is required for transport activity; thus DEPC probably inhibits NaDC-1 function by hindrance of the mobility of the carrier. We conclude that histidine residues are not critical for transport function in NaDC-1, although His-106 might be involved in determining protein expression or stability in the membrane.
Bai,
Expression cloning of NaDC-2, an intestinal Na(+)- or Li(+)-dependent dicarboxylate transporter.
1997, Pubmed,
Xenbase
Bai,
Expression cloning of NaDC-2, an intestinal Na(+)- or Li(+)-dependent dicarboxylate transporter.
1997,
Pubmed
,
Xenbase
Bindslev,
Histidyl residues at the active site of the Na/succinate co-transporter in rabbit renal brush borders.
1984,
Pubmed
Garrison,
The reaction between diethylpyrocarbonate and sulfhydryl groups in carbocylate buffers.
1975,
Pubmed
Geering,
A role for the beta-subunit in the expression of functional Na+-K+-ATPase in Xenopus oocytes.
1989,
Pubmed
,
Xenbase
Gerchman,
Histidine-226 is part of the pH sensor of NhaA, a Na+/H+ antiporter in Escherichia coli.
1993,
Pubmed
Hamm,
Renal handling of citrate.
1990,
Pubmed
Hirayama,
Glycosylation of the rabbit intestinal brush border Na+/glucose cotransporter.
1992,
Pubmed
Kaback,
A molecular mechanism for energy coupling in a membrane transport protein, the lactose permease of Escherichia coli.
1997,
Pubmed
Kunkel,
Rapid and efficient site-specific mutagenesis without phenotypic selection.
1985,
Pubmed
Maley,
Characterization of glycoproteins and their associated oligosaccharides through the use of endoglycosidases.
1989,
Pubmed
Markovich,
Expression cloning of rat renal Na+/SO4(2-) cotransport.
1993,
Pubmed
,
Xenbase
Miles,
Modification of histidyl residues in proteins by diethylpyrocarbonate.
1977,
Pubmed
Pajor,
Sequence and functional characterization of a renal sodium/dicarboxylate cotransporter.
1995,
Pubmed
,
Xenbase
Pajor,
Molecular cloning and functional expression of a sodium-dicarboxylate cotransporter from human kidney.
1996,
Pubmed
,
Xenbase
Pajor,
Functional differences between rabbit and human Na(+)-dicarboxylate cotransporters, NaDC-1 and hNaDC-1.
1996,
Pubmed
,
Xenbase
Pajor,
Characterization of the rabbit renal Na(+)-dicarboxylate cotransporter using antifusion protein antibodies.
1996,
Pubmed
,
Xenbase
Pourcher,
Melibiose permease of Escherichia coli: mutation of histidine-94 alters expression and stability rather than catalytic activity.
1992,
Pubmed
Püttner,
lac permease of Escherichia coli: histidine-205 and histidine-322 play different roles in lactose/H+ symport.
1986,
Pubmed
Sekler,
A cluster of cytoplasmic histidine residues specifies pH dependence of the AE2 plasma membrane anion exchanger.
1996,
Pubmed
Wang,
Mutational analysis of transmembrane histidines in the amiloride-sensitive Na+/H+ exchanger.
1995,
Pubmed
Wolffram,
Carrier-mediated transport is involved in mucosal succinate uptake by rat large intestine.
1994,
Pubmed
Wright,
Succinate and citrate transport in renal basolateral and brush-border membranes.
1987,
Pubmed
Wright,
Interactions between lithium and renal transport of Krebs cycle intermediates.
1982,
Pubmed
Wright,
Kinetics of sodium succinate cotransport across renal brush-border membranes.
1983,
Pubmed
