Transport of leucine, isoleucine and valine by luminal membrane vesicles from rabbit proximal tubule.
1. Transport of L- and D-isomers of leucine, isoleucine and valine by luminal membrane vesicles prepared from either the convoluted part (pars convoluta) or the straight part (pars recta) of rabbit proximal tubule was studied by a rapid filtration technique and by a spectrophotometric method using a potential-sensitive carbocyanine dye. 2. Both types of renal membrane vesicle take up the amino acids in a Na(+)-dependent, H(+)-independent and electrogenic manner. The L-isomers are transported with higher affinities than their corresponding D-forms, of which only D-leucine is taken up to a significant extent. 3. Membrane vesicles prepared from pars convoluta take up the L-amino acids by a single and common system. Filtration studies showed that the Km values for L-leucine and L-valine transport are, on average, 0.23 and 0.83 mM, respectively. The values of KA (the concentration of amino acid producing a half-maximal optical response) are comparable to those of Km, namely 0.18 mM for L-leucine and 0.60 mM for L-valine. KA for L-isoleucine transport was found to be 0.19 mM. D-Leucine is taken up by the same system but with a much lower affinity (KA = 7.2 mM). 4. Membrane vesicles prepared from pars recta possess two, and probably common, transport systems for the L-isomers of the amino acids. The average Michaelis-Menten constants were as follows: L-leucine, K1m = 0.17 mM, K2m = 6.5 mM; L-valine, K1m = 0.19 mM, K2m = 11.5 mM. The KA values were: L-leucine, K1A = 0.12 mM, K2A = 7.4 mM; L-valine, K1A = 0.18 mM, K2A = 10.0 mM; L-isoleucine, K1A = 0.17 mM, K2A = 9.0 mM. D-Leucine is taken up by a low-affinity system only (KA = 6.5 mM), which seems to be the same as the low-affinity system transporting the L-forms of the amino acids.
Jørgensen
K E
KE
Institute of Medical Biochemistry, University of Aarhus, Denmark.
Kragh-Hansen
U
U
Sheikh
M I
MI
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002232:Carbocyanines; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D007532:Isoleucine / Q000493:pharmacokinetics*; D007536:Isomerism; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D007930:Leucine / Q000493:pharmacokinetics*; D008297:Male; D011189:Potassium Chloride / Q000494:pharmacology; D011817:Rabbits; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors; D014633:Valine / Q000493:pharmacokinetics*
D000596:Amino Acids; D002232:Carbocyanines; D005456:Fluorescent Dyes; D007532:Isoleucine; C018919:3,3'-diethyloxadicarbocyanine; D012965:Sodium Chloride; D011189:Potassium Chloride; D007930:Leucine; D014633:Valine