Premium
Isolation and Characterization of Two Fatty Acid Binding Proteins from Mouse Brain
Author(s) -
MyersPayne Sean C.,
Hubbell Timothy,
Pu Lixia,
Schnütgen Frank,
Börchers Torsten,
Wood W. Gibson,
Spener Friedrich,
Schroeder Friedhelm
Publication year - 1996
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1996.66041648.x
Subject(s) - fatty acid binding protein , recombinant dna , biochemistry , fatty acid , heart type fatty acid binding protein , stearic acid , antiserum , cytosol , tricine , gel electrophoresis , myristic acid , free fatty acid receptor , biology , microbiology and biotechnology , chemistry , antibody , palmitic acid , enzyme , gene , polyunsaturated fatty acid , organic chemistry , immunology
Two fatty acid binding proteins (FABPs) were isolated from Swiss Webster mouse brains. Neither protein cross‐reacted with antisera to recombinant liver L‐FABP. One protein, designated brain H‐FABP, migrated on tricine sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) as a single band at 14.5 kDa with pl 4.9. Brain H‐FABP bound NBD‐stearic acid and cis ‐parinaric acid with K D values near 0.02 and 0.5 µ M , respectively. Brain H‐FABP cross‐reacted with affinity‐purified antisera to recombinant heart H‐FABP. The second protein, mouse brain B‐FABP, migrated on tricine SDS‐PAGE gels as a doublet at 16.0 and 15.5 kDa with pl values of 4.5 and 4.7, respectively. Brain B‐FABP bound NBD‐stearic acid and cis ‐parinaric acid with K D values near 0.01 and 0.7 µ M , respectively. The brain B‐FABP doublet was immunoreactive with affinity‐purified antibodies against recombinant mouse brain B‐FABP, but not with affinity‐purified antibodies against heart H‐FABP. [ 3 H]Oleate competition binding indicated that the two brain FABPs had distinct ligand binding specificities. Both bound fatty acids, fatty acyl CoA, and lysophosphatidic acid. Although both preferentially bound unsaturated fatty acids, twofold differences in specific saturated fatty acid binding were observed. Brain B‐FABP and brain H‐FABP represented 0.1 and 0.01% of brain total cytosolic protein, respectively. In summary, mouse brain contains two native fatty acid binding proteins, brain H‐FABP and brain B‐FABP.