Substitution of alanine for serine 250 in the murine fatty acid transport protein inhibits long chain fatty acid transport

S M Stuhlsatz-Krouper; N E Bennett; J E Schaffer

doi:10.1074/jbc.273.44.28642

Substitution of alanine for serine 250 in the murine fatty acid transport protein inhibits long chain fatty acid transport

J Biol Chem. 1998 Oct 30;273(44):28642-50. doi: 10.1074/jbc.273.44.28642.

Authors

S M Stuhlsatz-Krouper¹, N E Bennett, J E Schaffer

Affiliation

¹ Center for Cardiovascular Research, Department of Internal Medicine and the Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110-1010, USA.

PMID: 9786857
DOI: 10.1074/jbc.273.44.28642

Abstract

The murine fatty acid transport protein (FATP) was identified on the basis of its ability to facilitate uptake of long chain fatty acids (LCFAs) when expressed in mammalian cells. To delineate FATP domains important for transport function, we cloned the human heart FATP ortholog. Comparison of the human, murine, and yeast amino acid sequences identified a highly conserved motif, IYTSGTTGXPK, also found in a number of proteins that form adenylated intermediates. We demonstrate that depletion of intracellular ATP dramatically reduces FATP-mediated LCFA uptake. Furthermore, wild-type FATP specifically binds [alpha-32P]azido-ATP. Introduction of a serine to alanine substitution (S250A) in the IYTSGTTGXPK motif produces an appropriately expressed and metabolized mutant FATP that demonstrates diminished LCFA transport function and decreased [alpha-32P]azido-ATP binding. These results are consistent with a mechanism of action for FATP involving ATP binding that is dependent on serine 250 of the IYTSGTTGXPK motif.

MeSH terms

3T3 Cells
Adenosine Triphosphate / analogs & derivatives
Adenosine Triphosphate / metabolism
Alanine / chemistry*
Amino Acid Sequence
Amino Acid Substitution
Animals
Azides / metabolism
Biological Transport
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / pharmacology
Carrier Proteins / chemistry
Carrier Proteins / genetics
Carrier Proteins / metabolism*
Cloning, Molecular
Cyanides / pharmacology
DNA, Complementary
Fatty Acid Transport Proteins
Fatty Acids / metabolism*
Membrane Proteins / chemistry
Membrane Proteins / genetics
Membrane Proteins / metabolism*
Membrane Transport Proteins*
Mice
Molecular Sequence Data
Mutagenesis, Site-Directed
Protein Binding
Sequence Homology, Amino Acid
Serine / chemistry*

Substances

Azides
Carrier Proteins
Cyanides
DNA, Complementary
Fatty Acid Transport Proteins
Fatty Acids
Membrane Proteins
Membrane Transport Proteins
Slc27a4 protein, mouse
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Serine
2-azidoadenosine 5'-triphosphate
Adenosine Triphosphate
Alanine