An artificially evolved albumin binding module facilitates chemical shift epitope mapping of GA domain interactions with phylogenetically diverse albumins.

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TitleAn artificially evolved albumin binding module facilitates chemical shift epitope mapping of GA domain interactions with phylogenetically diverse albumins.
Publication TypeJournal Article
Year of Publication2007
AuthorsHe, Y, Chen, Y, Rozak, DA, Bryan, PN, Orban, J
JournalProtein Sci
Volume16
Issue7
Pagination1490-4
Date Published2007 Jul
ISSN0961-8368
KeywordsAmino Acid Sequence, Animals, Bacterial Proteins, Humans, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Sequence Data, Nerve Tissue Proteins, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Rabbits, Sequence Homology, Amino Acid, Serum Albumin
Abstract

Protein G-related albumin-binding (GA) modules occur on the surface of numerous Gram-positive bacterial pathogens and their presence may promote bacterial growth and virulence in mammalian hosts. We recently used phage display selection to evolve a GA domain, PSD-1 (phage selected domain-1), which tightly bound phylogenetically diverse albumins. With respect to PSD-1's broad albumin binding specificity, it remained unclear how the evolved binding epitope compared to those of naturally occurring GA domains and whether PSD-1's binding mode was the same for different albumins. We investigate these questions here using chemical shift perturbation measurements of PSD-1 with rabbit serum albumin (RSA) and human serum albumin (HSA) and put the results in the context of previous work on structure and dynamics of GA domains. Combined, these data provide insights into the requirements for broad binding specificity in GA-albumin interactions. Moreover, we note that using the phage-optimized PSD-1 protein significantly diminishes the effects of exchange broadening at the binding interface between GA modules and albumin, presumably through stabilization of a ligand-bound conformation. The employment of artificially evolved domains may be generally useful in NMR structural studies of other protein-protein complexes.

DOI10.1110/ps.072799507
Alternate JournalProtein Sci.
PubMed ID17567743
PubMed Central IDPMC2206689
Grant List1S10RR15744 / RR / NCRR NIH HHS / United States
GM62154 / GM / NIGMS NIH HHS / United States