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Related Experiment Videos

Renal organic cation and nucleoside transport.

Rong Chen1, Johan W Jonker, J Arly Nelson

  • 1Division of Pediatrics and Department of Experimental Therapeutics, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030, USA.

Biochemical Pharmacology
|July 19, 2002
PubMed
Summary

Organic cation transporter 1 (OCT1) transports deoxytubercidin (dTub), but OCT1 is not essential for its renal secretion in mice. This study clarifies dTub transport mechanisms and OCT1

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Area of Science:

  • Pharmacology
  • Molecular Biology
  • Biochemistry

Background:

  • Rat organic cation transporter 1 (rOCT1) transports the nucleoside analog deoxytubercidin (dTub).
  • rOCT1 transports the cationic form of dTub (dTub(+)), but rOCT2 does not transport dTub at pH 7.4.

Purpose of the Study:

  • To investigate the kinetic parameters (K(m), V(max)) of dTub(+) uptake by rOCT1 and rOCT2 at reduced pH (5.4).
  • To identify the regions responsible for differential dTub binding between rOCT1 and rOCT2.
  • To evaluate the role of OCT1 in the renal secretion of dTub using OCT1 knockout mice.

Main Methods:

  • Measurement of K(m) and V(max) for dTub(+) uptake in rOCT1 and rOCT2 at pH 5.4.
  • Construction and analysis of rOCT1/rOCT2 chimeras to map dTub binding domains.

Related Experiment Videos

  • Assessment of dTub tissue distribution and urinary excretion in OCT1 knockout and wild-type mice.

    • Main Results:

    • rOCT1 exhibits significantly higher transport efficiency (V(max)/K(m)) for dTub(+) than rOCT2 at pH 5.4.
    • The difference in dTub binding between rOCT1 and rOCT2 is localized to transmembrane domains 2-7.
    • OCT1 knockout mice showed no significant differences in renal elimination, plasma levels, or tissue distribution of dTub compared to wild-type mice.

    Conclusions:

    • dTub is a substrate for OCT1, with transport efficiency being significantly lower for OCT2 due to reduced affinity.
    • Transmembrane domains 2-7 of OCT1 and OCT2 are critical for dTub binding.
    • Despite being a substrate for OCT1, OCT1 is not essential for the renal secretion of dTub.