Multiple components of arginine and phenylalanine transport induced in neutral and basic amino acid transporter-cRNA-injected Xenopus oocytes.
The induced uptakes of L-[3H]phenylalanine and L-[3H]arginine in oocytes injected with clonal NBAT (neutral and basic amino acid transporter) cRNA show differential inactivation by pretreatment with N-ethylmaleimide (NEM), revealing at least two distinct transport processes. NEM-resistant arginine transport is inhibited by leucine and phenylalanine but not by alanine or valine; mutual competitive inhibition of NEM-resistant uptake of arginine and phenylalanine indicates that the two amino acids share a single transporter. NEM-sensitive arginine transport is inhibited by leucine, phenylalanine, alanine and valine. At least two NEM-sensitive transporters may be expressed because we have been unable to confirm mutual competitive inhibition between arginine and phenylalanine transport. The NEM-resistant transport mechanism appears to involve distinct but overlapping binding sites for cationic and zwitterionic substrates. NBAT is known to form oligomeric protein complexes in cell membranes, and its functional roles when expressed in Xenopus oocytes may include interaction with oocyte proteins, leading to increased native amino acid transport activities; these resemble NBAT-expressed activities in terms of NEM-sensitivity and apparent substrate range (including an unusual inhibition by beta-phenylalanine.
Peter
G J
GJ
Department of Anatomy and Physiology, University of Dundee, Scotland, U.K.
Davidson
I G
IG
Ahmed
A
A
McIlroy
L
L
Forrester
A R
AR
Taylor
P M
PM
D026923:Amino Acid Transport Systems, Basic; D026921:Amino Acid Transport Systems, Neutral; D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D001665:Binding Sites; D001693:Biological Transport, Active / Q000187:drug effects; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D007700:Kinetics; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism*; D018075:RNA, Complementary / Q000008:administration & dosage / Q000235:genetics*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014982:Xenopus laevis
D026923:Amino Acid Transport Systems, Basic; D026921:Amino Acid Transport Systems, Neutral; D000596:Amino Acids; D002352:Carrier Proteins; D018075:RNA, Complementary; D013439:Sulfhydryl Reagents; D010649:Phenylalanine; D001120:Arginine; D005033:Ethylmaleimide