LFABPLiver Fatty Acid Binding Protein
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Levels of urinary LFABP were elevated in normoalbuminuric patients than in the controls and were further increased with increasing levels of albuminuria, indicating its value in accurately reflecting severity of tubular damage in the early stage of DN [48].
Heemann et al., "Urinary LFABP and anaemia: distinct roles of urinary markers in type 2 diabetes," European Journal of Clinical Investigation, vol.
The binding constants of NBD stearate to L-FABP and SCP-2 were obtained by titrating a 2 mL sample of LFABP (25 nM) or SCP-2 (25 nM) in 10 mM phosphate buffer (pH 7.4) with small increments of NBD stearate at 24[degrees]C.
Displacement of NBD-stearate from L-FABP and SCP-2 was determined by incubating LFABP or SCP-2 (25 nM in 10 mM phosphate buffer) with NBD stearate (40 nM) for 12 min to obtain maximal fluorescence, followed by titration with increasing amount of ligand.
LFABP (-/-) null mice were obtained by targeted disruption of the L-FABP gene [36] and backcrossed to C57Bl/6N background to the N10 (99.9% homogeneity) generation.
C75 displaced ANS from LFABP with a Kt of 5.59 [+ or -] 0.31 [micro]M, thereby confirming weaker binding than observed with the NBD-stearic acid displacement assay.
Taken together, these data suggested that TOFyl-CoA did not actually displace NBD-stearic acid from L-FABP, but by binding to L-FABP the TOFyl-CoA instead shifted NBDstearic acid to a more hydrophobic environment within the LFABP binding pocket.
On the contrary, at 20 mM glucose and in the presence of TOFA the loss of LFABP decreased by nearly 50% the PPAR[alpha] transcription of CPT1A (Figure 12(b), open bars), CPT2 (Figure 12(d), open bars), or ACOX1 (Figure 12(f), open bars).
Regardless, however, it is important to note that while LFABP's affinity for C75 and C75-CoA was significantly lower than that of TOFA and its thioesters, nevertheless these affinities were in the range of the less potent fibrate activators of PPAR[alpha] such as bezafibrate and clofibrate [30, 31, 33, 69].
Third, L-FABP not only binds the fatty acid synthesis inhibitors, but several (tOfA, C75-CoA) also altered LFABP's secondary structure, suggesting that this in turn may facilitate L-FABP ligand signaling to PPAR[alpha].