L-Leucine Transport in Isolated Protoplasts of Vinca Suspension Culture: Characterization of Uptake |
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Authors: | Suzuki Masahiko |
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Institution: | Department of Pure and Applied Sciences, University of Tokyo 3-8-1 Komaba, Meguro-ku, Tokyo 153, Japan |
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Abstract: | The uptake of L-leucine into Vinca protoplasts was studied undervarious conditions. The uptake was highly pH-dependent, withthe optimal pH between 3.0 and 4.0. The uptake was also energydependent, since azide, 2,4-dinitrophenol (DNP), carbonyl cyanidem-chlorophenyl hydrazone (CCCP), and iodoacetate inhibited theuptake. Oligomycin, N,N'-dicycIohexyI carbodiimide (DCCD) andvanadate, but not ouabain, inhibited the uptake, suggestingthat ATPase for H+ electrogenic extrusion was necessary to theuptake of L-leucine. The uptake showed stereospecificity, butwas partially inhibited by other L-amino acids. A kinetic studyof the uptake showed that the uptake was multiphasic with threesaturable phases and one unsaturable phase which occurred atconcentrations of L-leucine over 1 mM. The Km values of thethree affinity sites were 1.4 x 103 M, 1.3 x 104M, 4.3 x 105 M; the maximum velocity values were 3.3x 108, 4.5 x 109, 1.8 x 109 mol/10 min/4x 106 cells. (Received April 18, 1981; Accepted August 25, 1981) |
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