Date of Award

1-1-2013

Language

English

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Chemistry

Content Description

1 online resource (iv, 48 pages) : illustrations (some color)

Dissertation/Thesis Chair

Alexander Shekhtman

Committee Members

Igor Lednev

Keywords

actin, genetic engineering, motor proteins, mutagenesis, nanotechnology, Adenosine triphosphate, Microfilament proteins, Mutagenesis

Subject Categories

Biochemistry | Molecular Biology

Abstract

Molecular motors are a remarkable feature within living organisms that are responsible for directional mechanical motion, which is driven by adenosine triphosphate (ATP) hydrolysis. Actin-binding molecular motors are of specific interest in the field of nanotechnology as filamentous actin is capable of carrying cargo, such as quantum dots, while it is translocated along a motor coated surface. The binding regions of motor proteins, which are known to interact with actin, such as Myosin, have been thoroughly examined and identified. Rapid genetic engineering of the ATP-hydrolyzing enzyme, adenosine kinase, to incorporate these binding regions is possible through the use of site- directed mutagenesis. The sequences, which were mutated into the ADKwt gene, were incorporated in an unstructured loop region. During the phosphate transfer, the mutants switch between open and closed conformational states. The binding affinity of the sequences to the actin is altered during this conformational switch, thus causing the motor to move along actin filament. The ADK mutants and their interaction with filamentous actin was monitored by an in vitro motility assay. Two different mutants of ADK were found to have retained enzymatic functionality after the mutagenesis as well as function as actin-based motor proteins.

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