Combining ultracentrifugation with fluorescence to follow the unfolding of modules 16-17 of complement receptor type 1

M D Kirkitadze, K Jumel, S E Harding, D T F Dryden, M Krych, J P Atkinson, P N Barlow

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Complement receptor type 1 (CR1) is a member of a family of regulators of complement activation with therapeutic potential. A fragment of CR1 comprising modules 16 and 17 was overexpressed as a recombinant nonglycosylated protein in Pichia pastoris. Intrinsic fluorescence studies of the unfolding of recombinant CR1 similar to 16-17 caused by increasing concentrations of guanidinium chloride revealed that the intermodular junction unfolds first, followed by module 17, with module 16 the last to unfold. Sedimentation velocity studies in the analytical ultracentrifuge revealed a corresponding clear change in conformation from the native macromolecules with an axial ratio of about 5:1 to a much more extended conformation.

Original languageEnglish
Title of host publicationANALYTICAL ULTRACENTRIFUGATION V
EditorsH Colfen
Place of PublicationBERLIN
PublisherSpringer-Verlag GmbH
Pages164-167
Number of pages4
ISBN (Print)3-540-66175-1
Publication statusPublished - 1999
Event11th Symposium on Analytical Ultracentrifugation - POTSDAM
Duration: 25 Mar 199926 Mar 1999

Conference

Conference11th Symposium on Analytical Ultracentrifugation
CityPOTSDAM
Period25/03/9926/03/99

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