Evolution of a de novo designed three-helix bundle protein: From structure to function and everything in between.

Format:

Book

Thesis/Dissertation

Author/Creator:

Walsh, Scott Thomas Russell.

Contributor:

University of Pennsylvania.

Language:

English

Subjects (All):

Biophysics.

Biochemistry.

0487.

0786.

Local Subjects:

0487.

0786.

Physical Description:

186 pages

Contained In:

Dissertation Abstracts International 61-06B.

System Details:

Mode of access: World Wide Web.

text file

Summary:

Proteins are natures' work-horses. Proteins are involved in every form of cell signaling and also carry out a vast amount of chemistry inside our bodies. Understanding how the protein's primary sequence encodes all the necessary information to fold into a unique three-dimensional structure remains unresolved. Designing proteins from scratch is an important endeavor in understanding the protein folding problem with implications for designing new functional proteins to do novel chemistry, as well as the treatment of human diseases such as cancer and allergic conditions. Here, a de novo designed three helix bundle protein, alpha3D, is studied. First the solution structure of alpha3D was determined using heteronuclear, multidimensional NMR methods and indeed shows that the structure of alpha3D folds into the desired topology. Second, the roles of thermodynamic stability and conformational specificity are explored by biophysical studies of hydrophobic core mutations into alpha3D. Increased thermodynamic stability of the core mutations into alpha3D potentially comes at the expense of conformational specificity. The backbone and side-chain dynamics of alpha 3D were also studied using NMR relaxation methods. At the backbone level, the picosecond-nanosecond motions of alpha3D are similar to natural proteins. The side-chain methyl groups in the hydrophobic core of alpha 3D show more dynamic ps-ns motions relative to natural proteins. Finally, the first steps into engineering function into alpha3D to inhibit a cell surface receptor in the progression of several allergic conditions has been accomplished using rational protein design and phage display methods.

Notes:

Source: Dissertation Abstracts International, Volume: 61-06, Section: B, page: 3053.

Adviser: William F. DeGrado.

Thesis (Ph.D.)--University of Pennsylvania, 2000.

Local Notes:

School code: 0175.

ISBN:

9780599822030

Access Restriction:

Restricted for use by site license.