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Backbone flexibility in protein design theory and experiment

Citation

Su, Alyce (1998) Backbone flexibility in protein design theory and experiment. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechTHESIS:05162011-132252955

Abstract

The role of backbone flexibility in protein design was studied. First, the effect of explicit backbone motion on the selection of amino acids in protein design was assessed in the core of the streptococcal protein Gβ1 domain (Gβ1). Concerted backbone motion was introduced by varying Gβ1's supersecondary structure parameter values. The stability and structural flexibility of seven of the redesigned proteins were determined experimentally. Core variants containing as many as six of ten possible mutations retained native-like properties. This result demonstrates that backbone flexibility can be combined with amino acid side-chain selection and that the selection algorithm is sufficiently robust to tolerate perturbations as large as 15% of the native parameter values. Second, a general, quantitative design method for computing de novo backbone templates was developed. The method had to compute atomic resolution backbones compatible with the atomistic sequence selection algorithm we were using and it had to be applicable to all protein motifs. We again developed a method that uses super-secondary structure parameters to determine the orientation among secondary structural elements, given a target protein fold. Possible backbone arrangements were screened using a cost function which evaluates core packing, hydrogen bonding, loop closure, and backbone torsional geometry. Given a specified number of residues for each secondary structural element, a family of optimal configurations was found. We chose three motifs to test our method (ββα, βαβ, and αα) since their combination could be used to approximate most possible backbone fold. The best structure found for the ββα motif is similar to a zinc finger, and the best structure for the ββα motif is similar to a segment of a β-barrel. The backbone obtained for the αα motif resembles minimized protein A. Last, our backbone design method was evaluated by testing the thermal stability and structural properties of the designed peptides using circular dichroism and 1D nuclear magnetic resonance. From these results, a set of heuristic rules was derived. Taken together, these studies suggest that de novo backbones assembled using our backbone design method may serve as adequate input templates for atomistic sequence selection algorithms.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:Physics
Degree Grantor:California Institute of Technology
Division:Physics, Mathematics and Astronomy
Major Option:Physics
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Mayo, Stephen L.
Thesis Committee:
  • Unknown, Unknown
Defense Date:18 May 1998
Record Number:CaltechTHESIS:05162011-132252955
Persistent URL:http://resolver.caltech.edu/CaltechTHESIS:05162011-132252955
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:6397
Collection:CaltechTHESIS
Deposited By: Benjamin Perez
Deposited On:16 May 2011 21:21
Last Modified:26 Dec 2012 04:35

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