| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
RESEARCH COMMUNICATION |
a Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona-08028, Spain
b Renal Division, Brigham and Women's Hospital, Harvard Institutes of Medicine, Boston, Massachusetts 02115, USA
c Departamento de Fisiología y Biofísica, Facultad de Medicina, Universidad de Chile, Casilla 70005, Santiago, Chile
4F2hc is an almost ubiquitous transmembrane protein in mammalian cells; upon expression in Xenopus laevis oocytes, it induces amino acid transport with characteristics of system y+L. Indirect evidence fostered speculation that function requires the association of 4F2hc with another protein endogenous to oocytes and native tissues. We show that expression of system y+L-like amino acid transport activity by 4F2hc in oocytes is limited by an endogenous factor and that direct covalent modification of external cysteine residue(s) of an oocyte membrane protein blocks system y+L/4F2hc transport activity, based on the following. 1) Induction of system y+L-like activity saturates at very low doses of human 4F2hc cRNA (0.1 ng/oocyte). This saturation occurs with very low expression of 4F2hc at the oocyte surface, and further increased expression of the protein at the cell surface does not result in higher induction of system y+L-like activity. 2) Human 4F2hc contains only two cysteine residues (C109 and C330). We mutated these residues, singly and in combination, to serine (C109S; CS1, C330S; CS2 and C109S-C330S, Cys-less). Mutation CS2 had no effect on the expressed system y+L-like transport activity, whereas C109S-containing mutants (CS1 and Cys-less) retained only partial y+L-like transport activity (30 to 50% of wild type). 3) Hg2+, the organic mercury compounds pCMB, and the membrane-impermeant pCMBS almost completely inactivated system y+L-like induced by human 4F2hc wild type and all the mutants studied. This was reversed by ß-mercaptoethanol, indicating that external cysteine residue(s) are the target of this inactivation. 4) Sensitivity to Hg2+ inactivation is increased by pretreatment of oocytes with ß-mercaptoethanol or in the C109S-containing mutants (CS1 and Cys-less). The increased Hg2+ reactivity of C109S-containing mutants supports the possibility that C109 may be linked by a disulfide bond to the Hg2+-targeted cysteine residue of the associated protein. These results indicate that 4F2hc is intimately associated with a membrane oocyte protein for the expression of system y+L amino acid transport activity. To our knowledge, this is the first direct evidence for a heteromultimeric protein structure of an organic solute carrier in mammals.—Estévez, R., Camps, M., Rojas, A. M., Testar, X., Devés, R., Hediger, M. A., Zorzano, A., Palacín, M. The amino acid transport system y+L/4F2hc is a heteromultimeric complex. FASEB J. 12, 1319–1329 (1998)
Key Words: oocyte • mutagenesis • erythrocyte • y+L transport activity • homologous protein
This article has been cited by other articles:
|
|
 |
|
  J. P. Laspiur, J. L. Burton, P. S. D. Weber, J. Moore, R. N. Kirkwood, and N. L. Trottier Dietary Protein Intake and Stage of Lactation Differentially Modulate Amino Acid Transporter mRNA Abundance in Porcine Mammary Tissue J. Nutr., September 1, 2009; 139(9): 1677 - 1684. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. R. Stam, E. G.J. Danchin, C. Rancurel, P. M. Coutinho, and B. Henrissat Dividing the large glycoside hydrolase family 13 into subfamilies: towards improved functional annotations of {alpha}-amylase-related proteins Protein Eng. Des. Sel., December 1, 2006; 19(12): 555 - 562. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Fernandez, M. Jimenez-Vidal, M. Calvo, A. Zorzano, F. Tebar, M. Palacin, and J. Chillaron The Structural and Functional Units of Heteromeric Amino Acid Transporters: THE HEAVY SUBUNIT rBAT DICTATES OLIGOMERIZATION OF THE HETEROMERIC AMINO ACID TRANSPORTERS J. Biol. Chem., September 8, 2006; 281(36): 26552 - 26561. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Cai, N. Bulus, P. M. Fonseca-Siesser, D. Chen, S. K. Hanks, A. Pozzi, and R. Zent CD98 modulates integrin {beta}1 function in polarized epithelial cells J. Cell Sci., March 1, 2005; 118(5): 889 - 899. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Soares-da-Silva and M. P. Serrao High- and low-affinity transport of L-leucine and L-DOPA by the hetero amino acid exchangers LAT1 and LAT2 in LLC-PK1 renal cells Am J Physiol Renal Physiol, August 1, 2004; 287(2): F252 - F261. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Jimenez-Vidal, E. Gasol, A. Zorzano, V. Nunes, M. Palacin, and J. Chillaron Thiol Modification of Cysteine 327 in the Eighth Transmembrane Domain of the Light Subunit xCT of the Heteromeric Cystine/Glutamate Antiporter Suggests Close Proximity to the Substrate Binding Site/Permeation Pathway J. Biol. Chem., March 19, 2004; 279(12): 11214 - 11221. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Blasco, J. J. Aramayona, A. I. Alcalde, J. Catalan, M. Sarasa, and V. Sorribas Rat kidney MAP17 induces cotransport of Na-mannose and Na-glucose in Xenopus laevis oocytes Am J Physiol Renal Physiol, October 1, 2003; 285(4): F799 - F810. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. E. Mann, D. L. Yudilevich, and L. Sobrevia Regulation of Amino Acid and Glucose Transporters in Endothelial and Smooth Muscle Cells Physiol Rev, January 1, 2003; 83(1): 183 - 252. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. V. Virkki, C. Franke, P. Somieski, and W. F. Boron Cloning and Functional Characterization of a Novel Aquaporin from Xenopus laevis Oocytes J. Biol. Chem., October 18, 2002; 277(43): 40610 - 40616. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Fernandez, M. Carrascal, F. Rousaud, J. Abian, A. Zorzano, M. Palacin, and J. Chillaron rBAT-b0,+AT heterodimer is the main apical reabsorption system for cystine in the kidney Am J Physiol Renal Physiol, September 1, 2002; 283(3): F540 - F548. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Chillaron, R. Roca, A. Valencia, A. Zorzano, and M. Palacin Heteromeric amino acid transporters: biochemistry, genetics, and physiology Am J Physiol Renal Physiol, December 1, 2001; 281(6): F995 - F1018. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. A. Wagner, F. Lang, and S. Broer Function and structure of heterodimeric amino acid transporters Am J Physiol Cell Physiol, October 1, 2001; 281(4): C1077 - C1093. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. L. Jack, I. T. Paulsen, and M. H. Saier The amino acid/polyamine/organocation (APC) superfamily of transporters specific for amino acids, polyamines and organocations Microbiology, August 1, 2000; 146(8): 1797 - 1814. [Abstract] [Full Text]
|
|
|
|
 |
|
 C. A Wagner, A. Broer, A. Albers, N. Gamper, F. Lang, and S. Broer The heterodimeric amino acid transporter 4F2hc/LAT1 is associated in Xenopus oocytes with a non-selective cation channel that is regulated by the serine/threonine kinase sgk-1 J. Physiol., July 1, 2000; 526(1): 35 - 46. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. H. Saier Jr. A Functional-Phylogenetic Classification System for Transmembrane Solute Transporters Microbiol. Mol. Biol. Rev., June 1, 2000; 64(2): 354 - 411. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. T.-Y. Ayuk, C. P. Sibley, P. Donnai, S. D'Souza, and J. D. Glazier Development and polarization of cationic amino acid transporters and regulators in the human placenta Am J Physiol Cell Physiol, June 1, 2000; 278(6): C1162 - C1171. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Zent, C. A. Fenczik, D. A. Calderwood, S. Liu, M. Dellos, and M. H. Ginsberg Class- and Splice Variant-specific Association of CD98 with Integrin beta Cytoplasmic Domains J. Biol. Chem., February 18, 2000; 275(7): 5059 - 5064. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Pfeiffer, J. Loffing, G. Rossier, C. Bauch, C. Meier, T. Eggermann, D. Loffing-Cueni, L. C. Kühn, and F. Verrey Luminal Heterodimeric Amino Acid Transporter Defective in Cystinuria Mol. Biol. Cell, December 1, 1999; 10(12): 4135 - 4147. [Abstract] [Full Text]
|
|
|
|
|
|
M. Pineda, E. Fernandez, D. Torrents, R. Estevez, C. Lopez, M. Camps, J. Lloberas, A. Zorzano, and M. Palacin Identification of a Membrane Protein, LAT-2, That Co-expresses with 4F2 Heavy Chain, an L-type Amino Acid Transport Activity with Broad Specificity for Small and Large Zwitterionic Amino Acids J. Biol. Chem., July 9, 1999; 274(28): 19738 - 19744. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Sato, M. Tamba, T. Ishii, and S. Bannai Cloning and Expression of a Plasma Membrane Cystine/Glutamate Exchange Transporter Composed of Two Distinct Proteins J. Biol. Chem., April 23, 1999; 274(17): 11455 - 11458. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D Markovich and R. Regeer Expression of membrane transporters in cane toad Bufo marinus oocytes J. Exp. Biol., January 8, 1999; 202(16): 2217 - 2223. [Abstract] [PDF]
|
|
|
|
|
|
D. Torrents, R. Estevez, M. Pineda, E. Fernandez, J. Lloberas, Y.-B. Shi, A. Zorzano, and M. Palacin Identification and Characterization of a Membrane Protein (y+L Amino Acid Transporter-1) That Associates with 4F2hc to Encode the Amino Acid Transport Activity y+L. A CANDIDATE GENE FOR LYSINURIC PROTEIN INTOLERANCE J. Biol. Chem., December 4, 1998; 273(49): 32437 - 32445. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Fenczik, R. Zent, M. Dellos, D. A. Calderwood, J. Satriano, C. Kelly, and M. H. Ginsberg Distinct Domains of CD98hc Regulate Integrins and Amino Acid Transport J. Biol. Chem., March 16, 2001; 276(12): 8746 - 8752. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
