de Koning, Harry P. and Jarvis, Simon M. (1997) Purine nucleobase transport in bloodstream forms of Trypanosoma brucei brucei is mediated by two novel transporters. Molecular and Biochemical Parasitology, 89 (2). pp. 245-258. ISSN 0166-6851. (The full text of this publication is not available from this repository)
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The mechanism and inhibitor sensitivity of hypoxanthine transport by bloodstream forms of Trypanosoma brucei brucei was investigated. The dose response curve for the inhibition of hypoxanthin transport (1 mu M) by guanosine was biphasic; approximate to 90% of transport activity was inhibited with a K-i value of 10.8 +/- 1.8 mu M, but 10% of the activity remained insensitive to concentrations as high as 2 mM. These two components of hypoxanthine transport are defined as guanosine-sensitive (H2) and guanosine-insensitive (H3). Hypoxanthine influx by both components was saturable, but there was a marked difference in their K-m values (123 +/- 15 nM and 4.7 +/- 0.9 mu M for H2 and H3, respectively) although the V-max values (1.1 +/- 0.2 and 1.1 +/- 0.1 pmol (10(7) cells)(-1) s(-1), n = 3) were similar. Hypoxanthine uptake via the H2 carrier was inhibited by purine bases and analogues as well as by some pyrimidine bases and one nucleoside (guanosine), whereas the H3 transporter was sensitive only to inhibition by purine nucleobases. H2-mediated hypoxanthine uptake was inhibited by ionophores, ion exchangers and the potential H+-ATPase inhibitors, N,N'-dicyclohexylcarbodiimide (DCCD) and N-ethylmalemide (NEM). Measurements of the intracellular pH and membrane potential of bloodstream trypanosomes in the presence and absence of these agents established a linear correlation between protonmotive force and rate of [H-3]hypoxanthine (30 nM) uptake. We conclude that hypoxanthine transport in bloodstream forms of T. b. brucei occurs by two transport systems with different affinities and substrate specificities, one of which, H2, appears to function as a H+/hypoxanthine symporter. (C) 1997 Elsevier Science B.V.
|Depositing User:||T. Nasir|
|Date Deposited:||24 Oct 2009 15:22|
|Last Modified:||20 Jun 2014 13:51|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/18388 (The current URI for this page, for reference purposes)|