Induced alignment and measurement of dipolar couplings of an SH2 domain through direct binding with filamentous phage
Authors: Dahlke Ojennus, D.; Mitton-Fry, R.M.; Wuttke, D.S.
Source: Journal of Biomolecular NMR, Volume 14, Number 2, June 1999 , pp. 175-179(5)
Abstract:Large residual ^15N-^1H dipolar couplings have been measured in a Src homology II domain aligned at Pf1 bacteriophage concentrations an order of magnitude lower than used for induction of a similar degree of alignment of nucleic acids and highly acidic proteins. An increase in ^1H and ^15N protein linewidths and a decrease in T_2 and T_1ρ relaxation time constants implicates a binding interaction between the protein and phage as the mechanism of alignment. However, the associated increased linewidth does not preclude the accurate measurement of large dipolar couplings in the aligned protein. A good correlation is observed between measured dipolar couplings and predicted values based on the high resolution NMR structure of the SH2 domain. The observation of binding-induced protein alignment promises to broaden the scope of alignment techniques by extending their applicability to proteins that are able to interact weakly with the alignment medium.
Document Type: Regular Paper
Affiliations: Department of Chemistry and Biochemistry, University of Colorado, Campus Box 215, Boulder, CO 80309-0215, U.S.A.
Publication date: June 1999