title: Binding and release of iron by gel-encapsulated human transferrin: evidence for a conformational search.
abstract: Human transferrin is a single-chain bilobal protein with each of the two similar but not identical lobes in turn composed of two domains. Each lobe may assume one of two stable structural conformations, open or closed, determined by a rigid rotation of the domains with respect to each other. In solution, the transformation of a lobe between open and closed conformations is associated with the release or binding of an Fe(III) ion. The results of the present study indicate that encapsulation of transferrin within a porous sol-gel matrix allows for a dramatic expansion, to days or weeks, of this interconversion time period, thus providing an opportunity to probe heretofore inaccessible transient intermediates. Sol-gel-encapsulated iron-free transferrin samples are prepared by using two protocols. In the first protocol, the equilibrium form of apotransferrin is encapsulated in the sol-gel matrix, whereas in the second protocol holotransferrin is first encapsulated and then iron is removed from the protein. Results of kinetic and spectroscopic studies allow for distinguishing between two models for iron binding. In the first, iron is assumed to bind to amino acid ligands of one domain, inducing a rigid rotation of the second domain to effect closure of the interdomain cleft. In the second, iron undertakes a conformational search among the thermally accessible states of the lobe, "choosing" the state which most nearly approximates the stable closed state when iron is bound. Our experimental results support the second mechanism.
authors.affiliation: Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Mahantesh S
authors.initials: MS
authors.lastname: Navati
authors.forename: Uri
authors.initials: U
authors.lastname: Samuni
authors.forename: Philip
authors.initials: P
authors.lastname: Aisen
authors.forename: Joel M
authors.initials: JM
authors.lastname: Friedman
mesh_terms: D001059:Apoproteins / Q000737:chemistry / Q000378:metabolism; D002214:Capsules; D005290:Ferric Compounds / Q000737:chemistry / Q000378:metabolism; D005782:Gels; D006801:Humans; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence / Q000379:methods; D013053:Spectrophotometry; D014168:Transferrin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001059:Apoproteins; D002214:Capsules; D005290:Ferric Compounds; D005782:Gels; D014168:Transferrin; C028284:apotransferrin; D007501:Iron