TY - JOUR
T1 - The light subunit of system bo,+ is fully functional in the absence of the heavy subunit
AU - Reig, Nùria
AU - Chillarón, Josep
AU - Bartoccioni, Paola
AU - Fernández, Esperanza
AU - Bendahan, Annie
AU - Zorzano, Antonio
AU - Kanner, Baruch
AU - Palacín, Manuel
AU - Bertran, Joan
PY - 2002/9/16
Y1 - 2002/9/16
N2 - The heteromeric amino acid transporters are composed of a type II glycoprotein and a non-glycosylated polytopic membrane protein. System bo,+ exchanges dibasic for neutral amino acids. It is composed of rBAT and bo,+AT, the latter being the polytopic membrane subunit. Mutations in either of them cause malfunction of the system, leading to cystinuria. bo,+AT-reconstituted systems from HeLa or MDCK cells catalysed transport of arginine that was totally dependent on the presence of one of the bo,+ substrates inside the liposomes. rBAT was essential for the cell surface expression of bo,+AT, but it was not required for reconstituted bo,+AT transport activity. No system bo,+ transport was detected in liposomes derived from cells expressing rBAT alone. The reconstituted bo,+AT showed kinetic asymmetry. Expressing the cystinuria-specific mutant A354T of bo,+AT in HeLa cells together with rBAT resulted in defective arginine uptake in whole cells, which was paralleled by the reconstituted bo,+AT activity. Thus, subunit bo,+AT by itself is sufficient to catalyse transmembrane amino acid exchange. The polytopic subunits may also be the catalytic part in other heteromeric transporters.
AB - The heteromeric amino acid transporters are composed of a type II glycoprotein and a non-glycosylated polytopic membrane protein. System bo,+ exchanges dibasic for neutral amino acids. It is composed of rBAT and bo,+AT, the latter being the polytopic membrane subunit. Mutations in either of them cause malfunction of the system, leading to cystinuria. bo,+AT-reconstituted systems from HeLa or MDCK cells catalysed transport of arginine that was totally dependent on the presence of one of the bo,+ substrates inside the liposomes. rBAT was essential for the cell surface expression of bo,+AT, but it was not required for reconstituted bo,+AT transport activity. No system bo,+ transport was detected in liposomes derived from cells expressing rBAT alone. The reconstituted bo,+AT showed kinetic asymmetry. Expressing the cystinuria-specific mutant A354T of bo,+AT in HeLa cells together with rBAT resulted in defective arginine uptake in whole cells, which was paralleled by the reconstituted bo,+AT activity. Thus, subunit bo,+AT by itself is sufficient to catalyse transmembrane amino acid exchange. The polytopic subunits may also be the catalytic part in other heteromeric transporters.
KW - BAT reconstitution
KW - BAT-rBAT exchanger
KW - Cystinuria
KW - Reabsorption
KW - Transport asymmetry
UR - http://www.scopus.com/inward/record.url?scp=0037119974&partnerID=8YFLogxK
U2 - 10.1093/emboj/cdf500
DO - 10.1093/emboj/cdf500
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C2 - 12234930
AN - SCOPUS:0037119974
SN - 0261-4189
VL - 21
SP - 4906
EP - 4914
JO - EMBO Journal
JF - EMBO Journal
IS - 18
ER -