Date Approved

11-2016

Document Type

Dissertation

Degree Name

PhD in Cell & Molecular Biology

Department

Graduate School of Biomedical Sciences

College

School of Osteopathic Medicine

First Advisor

Venkateswar Venkataraman, PhD

Committee Member 1

Mikhail Anikin, PhD

Committee Member 2

Dmitry Temiakov, PhD

Committee Member 3

Robert Nagele, PhD

Committee Member 4

Eric Moss, PhD

Subject(s)

Neuronal Calcium-Sensor Proteins, Neurocalcin, Hippocalcin, Calcium-Binding Proteins

Disciplines

Cell Biology | Laboratory and Basic Science Research | Medicine and Health Sciences | Molecular and Cellular Neuroscience | Molecular Biology

Abstract

Many neuronal functions, including learning and memory are driven by changes in intracellular Ca2+concentrations. The Neuronal Calcium Sensor (NCS) family of proteins is responsible for mediating the response to calcium. They are typically comprised of 4 EF hands; of which EF 2, 3, and 4 bind calcium.

Hypothesis: NCS proteins carry out unique, non-overlapping functions, and that specific characteristics of the family can be mapped to precise regions of the proteins.

Results: The effect on the following properties were investigated primarily on two highly similar NCS proteins, Neurocalcin Delta (NCALD) and Hippocalcin (HPCA): (1) Response to calcium was determined through two independent methods: change in tryptophan fluorescence and migration on a native polyacrylamide gel (CIMSA); (2) size exclusion chromatography, (3) Calcium binding, (4) Guanylate Cyclase stimulations and (5) sequence analysis. These results lead to the conclusion that specific aa locations are hyper-variable and pivotal in determining how the proteins respond.

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