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17 August 1994 Design of ribonuclease T1 mutants with tryptophan-59 in noncrystallographic conformations
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Abstract
Minimum perturbation mapping reveals that five side chain conformations of tryptophan-59 can potentially pack in the hydrophobic core of ribonuclease T1. In the crystallographic structure tryptophan-59 is in the trans perpendicular conformation. The other four wells in the tryptophan-59 (chi) 1 x (chi) 2 torsion space minimum perturbation mapping are trans antiperpendicular, gauche+ antiperpendicular, gauche+ perpendicular, and guache- perpendicular. The point mutations V33I and V33L are predicted to stabilize the trans antiperpendicular over the perpendicular conformation by 1.3 and 0.6 kcal/mol, respectively. The three gauche conformations require the creation of a new pocket within the protein core. The Mutation F80G creates the pocket for the gauche+ antiperpendicular tryptophan-59 conformation. The additional mutation V78A makes a more generous pocket for this conformation. The three mutation A19G, V78G and F80L creates the pocket for the gauche+ perpendicular conformation. A slightly more generous mutation at residue 80 may be required to fully stabilize this conformation. It does not appear possible to create a pocket that fits the gauche- perpendicular conformation better than the other two gauche conformations. The expression of ribonuclease-T1 mutants with tryptophan-59 in a gauche conformation probably requires filling in the crystallographic tryptophan-59 pocket. The mutations A22F, V33F, or V67F are all promising possibilities for this purpose.
© (1994) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Christopher Haydock "Design of ribonuclease T1 mutants with tryptophan-59 in noncrystallographic conformations", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182717
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