Aminoglycoside binding sites in the inner ears of guinea pigs.
With [125I]gentamicin as radioligand, the presence of aminoglycoside binding sites and kinetics of gentamicin binding to homogenates of organs of Corti, vestibular maculae, livers, spleens, and hearts of guinea pigs were investigated. The effects of temperature, osmolarity, 2,4-dinitrophenol, and digitonin on gentamicin binding were assessed. The affinities of several aminoglycosides for binding sites were tested. Gentamicin bound to cochlear and vestibular structures in a rapid and saturable fashion at a single class of noninteracting binding sites with Kds of 1.2.10(-6) and 3.10(-7) M and maximal binding capacities of 1.3 nmol and 43 pmol/mg of protein, respectively. In the liver, spleen, and heart, binding remained low and appeared to be nonspecific. In the organ of Corti, gentamicin uptake was unaffected by alterations in temperature or medium osmolarity or by 2,4-dinitrophenol, indicating that the uptake represented binding and not active transport. Digitonin at 10 nM increased markedly the uptake at 37 and 4 degrees C, suggesting the presence of internal binding sites. Various aminoglycosides compete for a common binding site.
Tran Ba Huy
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Département de Physiologie, Institut National de la Santé et de la Recherche Médicale Unité 251, Faculté de Médecine Xavier Bichat, Paris, France.
Deffrennes
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D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D001665:Binding Sites; D001667:Binding, Competitive; D005839:Gentamicins; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D009925:Organ of Corti / Q000378:metabolism*; D011869:Radioligand Assay; D014722:Vestibule, Labyrinth / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D005839:Gentamicins