title: Dissection of multi-protein complexes using mass spectrometry: subunit interactions in transthyretin and retinol-binding protein complexes.
abstract: Complexes formed between transthyretin and retinol-binding protein prevent loss of retinol from the body through glomerular filtration. The interactions between these proteins have been examined by electrospray ionization combined with time-of-flight mass analysis. Conditions were found whereby complexes of these proteins, containing from four to six protein molecules with up to two ligands, are preserved in the gas phase. Analysis of the mass spectra of these multimeric species gives the overall stoichiometry of the protein subunits and provides estimates for solution dissociation constants of 1.9 +/- 1.0 x 10(-7) M for the first and 3.5 +/- 1.0 x 10(-5) M for the second retinol-binding protein molecule bound to a transthyretin tetramer. Dissociation of these protein assemblies within the gas phase of the mass spectrometer shows that each retinol-binding protein molecule interacts with three transthyretin molecules. Mass spectral analysis illustrates not only a correlation with solution behavior and crystallographic data of a closely related protein complex but also exemplifies a general method for analysis of multi-protein assemblies.
authors.affiliation: Oxford Centre for Molecular Sciences, New Chemistry Laboratory, United Kingdom.
authors.forename: A A
authors.initials: AA
authors.lastname: Rostom
authors.forename: M
authors.initials: M
authors.lastname: Sunde
authors.forename: S J
authors.initials: SJ
authors.lastname: Richardson
authors.forename: G
authors.initials: G
authors.lastname: Schreiber
authors.forename: S
authors.initials: S
authors.lastname: Jarvis
authors.forename: R
authors.initials: R
authors.lastname: Bateman
authors.forename: C M
authors.initials: CM
authors.lastname: Dobson
authors.forename: C V
authors.initials: CV
authors.lastname: Robinson
mesh_terms: D000818:Animals; D002645:Chickens; D006801:Humans; D008958:Models, Molecular; D010449:Peptide Mapping; D011228:Prealbumin / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D012177:Retinol-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods
chemical_list: D011228:Prealbumin; D012177:Retinol-Binding Proteins