Solution Structure of the Apocalmodulin/neuro-P Complex

The interaction of apocalmodulin with a peptide (Neurop ) derived from the calmodulin-binding domain of the neural-specific protein neuromodulin has been studied by nuclear magnetic resonance (NMR). This complex is in fast exchange with its components on the NMR timescale, and the dissociation constant for the complex is in the micromolar range. This thesis presents the solution structure of the apoCaM. Neurop complex. The 1H, 13C, and 15N resonance assignments for the calmodulin and Neurop components of the complex were determined using multidimensional, multinuclear and triple resonance NMR methods. NOE-derived distance restraints and angular restraints were likewise determined. A total of 1910 intraprotein, 108 intrapeptide, and 18 protein-peptide NOEs were used to generate a family of structures. In addition, 217 torsional restraints and 68 hydrogen bond restraints were used to refine the final generated structures. A family of 48 refined structures was chosen for analysis on the basis of lowest energy and violations. The atomic r. m. s. d. for this family was 1.3 A for main chain atoms and 1.78 A for all heavy atoms. The average structure differs from other known calmodulin-peptide structures in possessing protein domains in which the hydrophobic surfaces normally involved in protein-peptide interactions are either not present (N-terminal domain) or are reduced in size (C-terminal domain). The result is a complex in which the peptide is loosely held by a pair of salt links, one to each domain of the protein, and hydrophobic interactions with the C-terminal domain.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Short, James Howard

Contributors dc:contributor

• Wand, A. Joshua

• Chemistry, Biochemistry

Language dc:language

eng

Identifier

(MiAaPQ)AAI9912378

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/84899