Abstract
Purpose. Previously, we suggested that alkylglucoside can be aneffective vector for renal-specific drug delivery (Suzuki et al., J. Pharmacol.Exp. Ther., 288:57–61, 1999). The purpose of the present study is tocharacterize the membrane protein which is recognized by thisalkylglucoside.
Methods. The binding of [125I] tyrosine conjugated with aoctylthioglucoside (Glc-S-C8-[125I]Tyr) Glc-S-C8-[125I]Tyr to crude membranefractions of kidney was determined. In addition, the membrane wascross-linked with this alkylglucoside and examined by sodium dodecylsulfate-polyacrylamide gel electrophoresis.
Results. Glc-S-C8-[125I]Tyr was shown to have a specific binding siteon the kidney membrane (Kd = 931 nM and Bmax = 987pmol/mg protein). Cross-linking of the membrane with Glc-S-C8-[125I]Tyrresulted in the detection of a protein (Mr = 62,000), which wasunaffected by reducing agents. The results of this cross-linking study wereconsistent with previous information on its localization and bindingcharacteristics.
Conclusions. The kidney membrane protein, to which alkylglucosidebinds in a specific manner, has a molecular weight of 62,000.Cross-linking is a useful tool for detecting this novel membrane proteinin kidney.
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REFERENCES
K. Suzuki, T. Ando, H. Susaki, K. Mimori, S. Nakabayashi, and Y. Sugiyama. Structual requirements for alkylglycoside-type renal targeting vector. Pharm. Res. 16:1026–1034 (1999).
K. Suzuki, H. Susaki, S. Okuno, H. Yamada, H. K. Watanabe, and Y. Sugiyama. Specific renal delivery of sugar-modified low-molecular-weight peptides. J. Pharmacol. Exp. Ther. 288:888–897 (1999).
K. Suzuki, H. Susaki, S. Okuno, and Y. Sugiyama. Renal drug targeting using a vector “alkylglycoside.” J. Pharmacol. Exp. Ther. 288:57–64 (1999).
D. K. Meijer and P. van der Sluijs. Covalent and noncovalent protein binding of drugs: implications for hepatic clearance, storage, and cell-specific drug delivery. Pharm. Res. 6:105–118 (1989).
W. Sadee, V. Drubbisch, and G. L. Amidon. Biology of membrane transport proteins. Pharm. Res. 12:1823–1837 (1995).
H. Saito, S. Masuda, and K. Inui. Cloning and functional characterization of a novel rat organic anion transporter mediating baso-lateral uptake of methotrexate in the kidney. J. Biol. Chem. 271:20719–20725 (1996).
A. M. Pajor. Molecular cloning and functional expression of a sodium-dicarboxylate cotransporter from human kidney. Am. J. Physiol. 270:F642–648 (1996).
M. A. Hediger and D. B. Rhoads. Molecular physiology of sodium-glucose cotransporters. Physiol. Rev. 74:993–1026 (1994).
W. H. Hunter and F. C. Greenwood. Preparation of iodine-131 labeled human growth hormone of high specific activity. Nature 194:495–496 (1962).
F. L. Stassen, R. W. Erickson, W. F. Huffman, J. Stefankiewicz, L. Sulat, and V. D. Wiebelhaus. Molecular mechanisms of novel antagonists: analysis of the effects on vasopressin binding and adenylate cyclase activation in animal and human kidney. J. Pharmacol. Exp. Ther. 223:50–54 (1982).
S. M. Grassl and P. S. Aronson. Na+/HCO3-co-transport in basolateral membrane vesicles isolated from rabbit renal cortex. J. Biol. Chem. 261:8778–8783 (1986).
H. Walter. Tightness and orientation of vesicles from guinea-pig kidney estimated from reactions of adenosine triphosphatase dependent on sodium and potassium ions. Eur. J. Biochem. 58:595–601 (1975).
K. Yamaoka, Y. Tanigawara, T. Nakagawa, and T. Uno. A pharmacokinetic analysis program (multi) for microcomputer. J. Pharmacobio. Dyn. 4:879–885 (1981).
M. T. Vincenzini, T. Iantomasi, M. Stio, C. Treves, F. Favilli, and P. Vanni. 1-O-n-octyl-beta-D-glucopyranoside as a competitive inhibitor of Na+-dependent D-glucose cotransporter in the small intestine brush-border membrane. Biochim. Biophys. Acta 903:273–276 (1987).
H. Kipp, J. T. Lin, and R. K. Kinne. Interactions of alkylglucosides with the renal sodium/D-glucose cotransporter. Biochim. Biophys. Acta 1282:125–130 (1996).
W. S. Lee, Y. Kanai, R. G. Wells, and M. A. Hediger. The high affinity Na+/glucose cotransporter. Re-evaluation of function and distribution of expression. J. Biol. Chem. 269:12032–12039 (1994).
R. J. Turner and A. Moran. Heterogeneity of sodium-dependent D-glucose transport sites along the proximal tubule: evidence from vesicle studies. Am. J. Physiol. 242:F406–414 (1982).
R. J. Turner and A. Moran. Stoichiometric studies of the renal outer cortical brush border membrane D-glucose transporter. J. Membr. Biol. 67:73–80 (1982).
R. J. Turner and A. Moran. Further studies of proximal tubular brush border membrane D-glucose transport heterogeneity. J. Membr. Biol. 70:37–45 (1982).
H. Kipp, R. K. Kinne, and J. T. Lin. Synthesis of the photoaffinity label [1'-14C]-6C-(azimethyl)octylglucoside and its reaction with isolated renal brush border membranes. Anal. Biochem. 245:61–68 (1997).
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Watanabe, Y., Suzuki, H., Suzuki, K. et al. Detection of the Membrane Protein Recognized by the Kidney-Specific Alkylglucoside Vector. Pharm Res 17, 49–54 (2000). https://doi.org/10.1023/A:1007566408323
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DOI: https://doi.org/10.1023/A:1007566408323