Amino acid sequences of pyridoxal 5'-phosphate binding sites and fluorescence resonance energy transfer in chicken liver fatty acid synthase

Biochemistry ◽  
1989 ◽  
Vol 28 (9) ◽  
pp. 3781-3788 ◽  
Author(s):  
Soo Ik Chang ◽  
Gordon G. Hammes
Keyword(s):  
Energy Transfer ◽  
Fatty Acid ◽  
Amino Acid ◽  
Binding Sites ◽  
Fatty Acid Synthase ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Amino Acid Sequences ◽  
Liver Fatty Acid ◽  
Phosphate Binding

scholarly journals Physical Association between the Adipocyte Fatty Acid-binding Protein and Hormone-sensitive Lipase

2004 ◽  
Vol 279 (50) ◽  
pp. 52399-52405 ◽  
Author(s):  
Anne J. Smith ◽  
Mark A. Sanders ◽  
Brian R. Thompson ◽  
Constantine Londos ◽  
Fredric B. Kraemer ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fatty Acid ◽  
Fluorescence Resonance Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Hormone Sensitive Lipase ◽  
Fatty Acid Binding ◽  
Hormone Sensitive ◽  
Fluorescence Resonance

Previousin vitrostudies have established that hormone sensitive lipase (HSL) and adipocyte fatty acid-binding protein (AFABP) form a physical complex that presumably positions the FABP to accept a product fatty acid generated during catalysis. To assess AFABP-HSL interaction within a cellular context, we have used lipocytes derived from 293 cells (C8PA cells) and examined physical association using fluorescence resonance energy transfer. Transfection of C8PA cells with cyan fluorescent protein (CFP)-HSL, yellow fluorescent protein (YFP)-adipocyte FABP, or YFP-liver FABP revealed that under basal conditions each protein was cytoplasmic. In the presence of 20 μmforskolin, CFP-HSL translocated to the triacylglycerol droplet, coincident with BODIPY-FA labeled depots. Fluorescence resonance energy transfer analysis demonstrated that CFP-HSL associated with YFP-adipocyte FABP in both basal and forskolin-treated cells. In contrast, little if any fluorescence resonance energy transfer could be detected between CFP-HSL and YFP-liver FABP. These results suggest that a pre-lipolysis complex containing at least AFABP and HSL exists and that the complex translocates to the surface of the lipid droplet.

scholarly journals Measuring distances between TRPV1 and the plasma membrane using a noncanonical amino acid and transition metal ion FRET

2016 ◽  
Vol 147 (2) ◽  
pp. 201-216 ◽  
Author(s):  
William N. Zagotta ◽  
Moshe T. Gordon ◽  
Eric N. Senning ◽  
Mika A. Munari ◽  
Sharona E. Gordon
Keyword(s):  
Plasma Membrane ◽  
Energy Transfer ◽  
Amino Acid ◽  
Membrane Proteins ◽  
Transition Metal ◽  
Fluorescence Resonance Energy Transfer ◽  
Metal Ion ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Noncanonical Amino Acid

Despite recent advances, the structure and dynamics of membrane proteins in cell membranes remain elusive. We implemented transition metal ion fluorescence resonance energy transfer (tmFRET) to measure distances between sites on the N-terminal ankyrin repeat domains (ARDs) of the pain-transducing ion channel TRPV1 and the intracellular surface of the plasma membrane. To preserve the native context, we used unroofed cells, and to specifically label sites in TRPV1, we incorporated a fluorescent, noncanonical amino acid, L-ANAP. A metal chelating lipid was used to decorate the plasma membrane with high-density/high-affinity metal-binding sites. The fluorescence resonance energy transfer (FRET) efficiencies between L-ANAP in TRPV1 and Co2+ bound to the plasma membrane were consistent with the arrangement of the ARDs in recent cryoelectron microscopy structures of TRPV1. No change in tmFRET was observed with the TRPV1 agonist capsaicin. These results demonstrate the power of tmFRET for measuring structure and rearrangements of membrane proteins relative to the cell membrane.

Evaluation of a D-amino-acid-containing fluorescence resonance energy transfer peptide library for profiling prokaryotic proteases

2013 ◽  
Vol 441 (1) ◽  
pp. 38-43 ◽  
Author(s):  
Wendy E. Kaman ◽  
Ingrid Voskamp-Visser ◽  
Denise M.C. de Jongh ◽  
Hubert P. Endtz ◽  
Alex van Belkum ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Amino Acid ◽  
Fluorescence Resonance Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Peptide Library ◽  
Fluorescence Resonance
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