In situ kinetic characterization of methylthioadenosine transport by the adenosine transporter (P2) of the African Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense.
African trypanosomes are parasitic flagellates that live in the connective tissues of the host. Trypanosomes must obtain from their host adenine/adenosine and other nucleosides that can be salvaged through enzymatic cleavage. Methylthioadenosine (MTA) is a byproduct of polyamine metabolism, formed from the donation of an aminopropyl moiety by decarboxylated S-adenosylmethionine (dcAdoMet) to form spermidine. MTA is then cleaved phosphorolytically by MTA phosphorylase to methylthioribose-1-phosphate (MTR-1-P) and adenine. The uptake of MTA was compared with that of adenosine in two strains: Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense. The K(m) values for MTA and adenosine (with 5 mM inosine) transport by T. b. brucei were 1.4 and 0.175 mM, and the V(max) values were 70 and 7.8 micromol/L/min, respectively. The K(m) values for T. b. rhodesiense MTA and adenosine (with 5 mM inosine) transport were 1.2 and 0.11 mM, and the V(max) values were 52.6 and 2.9 micromol/L/min, respectively. Since MTA was not competitive with either AdoMet (100 microM), inosine (100 microM), or the methionine precursor ketomethylthiobutyrate (100 microM), it appears that MTA enters through the P(2) (adenosine/adenine) transport site. From this study and our previous work, we determined that these organisms transport adenylated intermediates of methionine metabolism found in sera for purine salvage and as an ancillary source of methionine. The significant ability of African trypanosomes to transport MTA and related intermediates is an important consideration in the design and development of selective chemotherapeutic agents.
Goldberg
B
B
Department of Biology, St. Francis College, 180 Remsen Street, Brooklyn Heights, 11201, Brooklyn, NY, USA. bg43@nyu.edu
Rattendi
D
D
Lloyd
D
D
Sufrin
J R
JR
Bacchi
C J
CJ
D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000493:pharmacokinetics*; D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D033703:Nucleoside Transport Proteins; D011073:Polyamines / Q000378:metabolism; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D013872:Thionucleosides / Q000378:metabolism / Q000493:pharmacokinetics*; D014346:Trypanosoma brucei brucei / Q000378:metabolism*; D016833:Trypanosoma brucei rhodesiense / Q000378:metabolism*
C117012:5'-deoxy-5'-(hydroxyethylthio)adenosine; D002352:Carrier Proteins; D008565:Membrane Proteins; D033703:Nucleoside Transport Proteins; D011073:Polyamines; D013872:Thionucleosides; C021714:2-methylthioadenosine; D012435:S-Adenosylhomocysteine; D008715:Methionine; D000241:Adenosine