Dissection of multi-protein complexes using mass spectrometry: Subunit interactions in transthyretin and retinol-binding protein complexes

✍ Scribed by Adam A. Rostom; Margaret Sunde; Samantha J. Richardson; Gerhard Schreiber; Stuart Jarvis; Robert Bateman; Christopher M. Dobson; Carol V. Robinson

• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 290 KB
• Volume: 33
• Category: Article
• ISSN: 0887-3585
• DOI: 10.1002/(sici)1097-0134(1998)33:2+<3::aid-prot2>3.0.co;2-h

✦ Synopsis

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-offlight 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 ؋ 10 ؊7 M for the first and 3.5 ؎ 1.0 ؋ 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. Proteins