Identification of two leucine-sensitive lysine transport activities in human placental basal membrane

Placenta. 1997 Nov;18(8):649-55. doi: 10.1016/s0143-4004(97)90006-0.

Abstract

The recent demonstration of multiple high-affinity leucine-sensitive cationic transport systems prompted this investigation of their role in lysine uptake in basal cell membrane. Transport of lysine by basal membrane was saturable at both 22 and 37 degrees C and linear in time to 1 min and 30 sec, respectively. At 22 degrees C, at least two systems were active. The portion of uptake inhibited by the sulphydryl binding reagent N-ethylmaleimide (NEM) but not by leucine in the absence of sodium had a high K(m) and high Vmax and was attributed to system y+. NEM-insensitive uptake was fitted by a one-system model with K(m) (+/- s.e.) of 4 +/- 1 microM and a Vmax of 0.9 +/- 0.1 pmol/mg protein/min. This component was completely inhibited by leucine in the absence of sodium but not by glutamine in the presence of sodium. Therefore, it was attributed to system bo,+. At 37 degrees C, at least three systems were active. For essentially the same reasons as above the NEM inhibitable uptake was attributed to system y+. NEM-insensitive uptake was fitted by a one-system model with K(m) of 26 +/- 7 microM and Vmax of 11.1 +/- 2.8 pmol/mg protein/30 sec. Inhibition studies, however, indicated its heterogeneity. NEM-insensitive saturable uptake was only partially inhibited by either leucine in the absence of sodium (system bo,+) or by glutamine in the presence of sodium (system y+L). It is concluded that the NEM-insensitive portion of lysine uptake at 37 degrees C represents activity of both system bo,+ and the temperature-sensitive system y+L. As a previous investigation indicates, only one of these (system y+L) is present in the more specialized microvillous membrane. The demonstration of functional differences in the high affinity leucine transporters of basal and microvillous membrane in this and our previous investigations suggest that the two membranes possess different transport or modifier proteins.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Transport Systems, Basic
  • Basement Membrane / drug effects
  • Basement Membrane / metabolism
  • Carrier Proteins / metabolism*
  • Chorionic Villi / drug effects
  • Chorionic Villi / metabolism*
  • Ethylmaleimide / pharmacology
  • Female
  • Glutamine / pharmacology
  • Humans
  • In Vitro Techniques
  • Kinetics
  • Leucine / pharmacology*
  • Lysine / metabolism*
  • Membrane Proteins / metabolism*
  • Pregnancy
  • Temperature
  • Trophoblasts / drug effects
  • Trophoblasts / metabolism*

Substances

  • Amino Acid Transport Systems, Basic
  • Carrier Proteins
  • Membrane Proteins
  • Glutamine
  • Leucine
  • Lysine
  • Ethylmaleimide