Granulocyte-macrophage colony-stimulating factor increases L-arginine transport through the induction of CAT2 in bone marrow-derived macrophages.
L-arginine transport is crucial for macrophage activation because it supplies substrate for the key enzymes nitric oxide synthase 2 and arginase I. These enzymes participate in classic and alternative activation of macrophages, respectively. Classic activation of macrophages is induced by type I cytokines, and alternative activation is induced by type II cytokines. The granulocyte macrophage colony-stimulating factor (GM-CSF), in addition to inducing proliferation and differentiation of macrophages, activates arginase I, but its action on L-arginine transport is unknown. We studied the L-arginine transporters that are active in mouse primary bone marrow-derived macrophages (BMM) and examined the effect of GM-CSF treatment on transport activities. Under basal conditions, L-arginine entered mainly through system y(+)L (>75%). The remaining transport was explained by system y(+) (<10%) and a diffusion component (10-15%). In response to GM-CSF treatment, transport activity increased mostly through system y(+) (>10-fold), accounting for about 40% of the total L-arginine transport. The increase in y(+) activity correlated with a rise in cationic amino acid transporter (CAT)-2 mRNA and protein. Furthermore, GM-CSF induced an increase in arginase activity and in the conversion of L-arginine to ornithine, citrulline, glutamate, proline, and polyamines. BMM obtained from CAT2-knockout mice responded to GM-CSF by increasing arginase activity and the expression of CAT1 mRNA, which also encodes system y(+) activity. Nonetheless, the increase in CAT1 activity only partially compensated the lack of CAT2 and L-arginine metabolism was hardly stimulated. We conclude that BMM present mainly y(+)L activity and that, in response to GM-CSF, l-arginine transport augments through CAT2, thereby increasing the availability of this amino acid to the cell.
Martín
Lorena
L
Department of Biochemistry and Molecular Biology, University of Barcelona, Avenida Diagonal 645, Barcelona E-08028, Spain.
Comalada
Mónica
M
Marti
Luc
L
Closs
Ellen I
EI
MacLeod
Carol L
CL
Martín del Río
Rafael
R
Zorzano
Antonio
A
Modolell
Manuel
M
Celada
Antonio
A
Palacín
Manuel
M
Bertran
Joan
J
D000818:Animals; D001120:Arginine / Q000378:metabolism*; D001693:Biological Transport, Active / Q000187:drug effects / Q000502:physiology; D001853:Bone Marrow / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D027202:Cationic Amino Acid Transporter 2 / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice
D027202:Cationic Amino Acid Transporter 2; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D001120:Arginine