Back to Search
Start Over
GLUT-4 NH2 terminus contains a phenylalanine-based targeting motif that regulates intracellular sequestration.
- Source :
-
The Journal of cell biology [J Cell Biol] 1993 Jun; Vol. 121 (6), pp. 1221-32. - Publication Year :
- 1993
-
Abstract
- Expression of chimeras, composed of portions of two different glucose transporter isoforms (GLUT-1 and GLUT-4), in CHO cells had indicated that the cytoplasmic NH2 terminus of GLUT-4 contains important targeting information that mediates intracellular sequestration of this isoform (Piper, R. C., C. Tai, J. W. Slot, C. S. Hahn, C. M. Rice, H. Huang, D. E. James. 1992. J. Cell Biol. 117:729-743). In the present studies, the amino acid constituents of the GLUT-4 NH2-terminal targeting domain have been identified. GLUT-4 constructs containing NH2-terminal deletions or alanine substitutions within the NH2 terminus were expressed in CHO cells using a Sindbis virus expression system. Deletion of eight amino acids from the GLUT-4 NH2 terminus or substituting alanine for phenylalanine at position 5 in GLUT-4 resulted in a marked accumulation of the transporter at the plasma membrane. Mutations at other amino acids surrounding Phe5 also caused increased cell surface expression of GLUT-4 but not to the same extent as the Phe5 mutation. GLUT-4 was also localized to clathrin lattices and this colocalization was abolished when either the first 13 amino acids were deleted or when Phe5 was changed to alanine. To ascertain whether the targeting information within the GLUT-4 NH2-terminal targeting domain could function independently of the glucose transporter structure this domain was inserted into the cytoplasmic tail of the H1 subunit of the asialoglycoprotein receptor. H1 with the GLUT-4 NH2 terminus was predominantly localized to an intracellular compartment similar to GLUT-4 and was sequestered more from the cell surface than was the wild-type H1 protein. It is concluded that the NH2 terminus of GLUT-4 contains a phenylalanine-based targeting motif that mediates intracellular sequestration at least in part by facilitating interaction of the transporter with endocytic machinery located at the cell surface.
- Subjects :
- Amino Acid Sequence
Animals
Biological Transport
CHO Cells
Clathrin
Cricetinae
Deoxyglucose metabolism
Glucose Transporter Type 4
Molecular Sequence Data
Mutation
Phenylalanine
Protein Sorting Signals physiology
Sequence Deletion
Structure-Activity Relationship
Subcellular Fractions metabolism
Monosaccharide Transport Proteins chemistry
Monosaccharide Transport Proteins metabolism
Muscle Proteins
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9525
- Volume :
- 121
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- The Journal of cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 8509445
- Full Text :
- https://doi.org/10.1083/jcb.121.6.1221