| Title: | Probing the structure of the ligand binding cavity of lipocalins by fluorescence spectroscopy |
| Author(s): | Patel RC; Lange D; McConathy WJ; Patel YC; Patel SC; |
| Address: | "Department of Chemistry, Clarkson University, Potsdam, NY 13699-5810, USA" |
| ISSN/ISBN: | 0269-2139 (Print) 0269-2139 (Linking) |
| Abstract: | "The lipocalin superfamily constitutes a phylogenetically conserved group of more than 40 proteins that function in the binding and transport of a variety of physiologically important ligands. Members of this family subserve diverse functions as carriers of retinoids (retinol binding protein), odorants (odorant binding proteins), chromophores (insecticyanin, INS), pheromones (aphrodisin) and sterols (apolipoprotein D, apoD). Despite the pivotal importance of the ligand binding function of these proteins, a suitable approach for characterizing the molecular determinants of such binding has not been available. In studies using three homogeneously purified lipocalins INS, beta-lactoglobulin (BLG) and human apoD, we find that the fluorescence reporter BIS (1,1'-bi(4-anilino) naphthalene-5,5'-disulfonic acid) is an ideal candidate for use in rapid kinetic experiments and in fluorescence resonance energy transfer (FRET). These methods require only small amounts of reagents and yield molecular coordinates of the ligand binding cavity of lipocalins in solution that are in remarkably close agreement to those obtained from crystallographic work with solids. Extremely fast ligand binding dynamics is indicated" |
| Keywords: | Animals Apolipoproteins/*chemistry/metabolism Apolipoproteins D Binding Sites Carrier Proteins/*chemistry/metabolism Female Hemolymph/chemistry Humans *Insect Proteins Invertebrate Hormones/*chemistry/metabolism Lactoglobulins/*chemistry/metabolism Ligand; |
| Notes: | "MedlinePatel, R C Lange, D McConathy, W J Patel, Y C Patel, S C eng NS 34339-03/NS/NINDS NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England 1997/06/01 Protein Eng. 1997 Jun; 10(6):621-5. doi: 10.1093/protein/10.6.621" |
