کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1959406 1057937 2007 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
FRET Study of Membrane Proteins: Determination of the Tilt and Orientation of the N-Terminal Domain of M13 Major Coat Protein
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
FRET Study of Membrane Proteins: Determination of the Tilt and Orientation of the N-Terminal Domain of M13 Major Coat Protein
چکیده انگلیسی

A formalism for membrane protein structure determination was developed. This method is based on steady-state FRET data and information about the position of the fluorescence maxima on site-directed fluorescent labeled proteins in combination with global data analysis utilizing simulation-based fitting. The methodology was applied to determine the structural properties of the N-terminal domain of the major coat protein from bacteriophage M13 reconstituted into unilamellar DOPC/DOPG (4:1 mol/mol) vesicles. For our purpose, the cysteine mutants A7C, A9C, N12C, S13C, Q15C, A16C, S17C, and A18C in the N-terminal domain of this protein were produced and specifically labeled with the fluorescence probe AEDANS. The energy transfer data from the natural Trp-26 to AEDANS were analyzed assuming a two-helix protein model. Furthermore, the polarity Stokes shift of the AEDANS fluorescence maxima is taken into account. As a result the orientation and tilt of the N-terminal protein domain with respect to the bilayer interface were obtained, showing for the first time, to our knowledge, an overall α-helical protein conformation from amino acid residues 12–46, close to the protein conformation in the intact phage.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: - Volume 92, Issue 4, 15 February 2007, Pages 1296–1305
نویسندگان
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