Crystal Structure Determination Of Two New Red Fluorescent Proteins By X-Ray Crystallography

Date
2017-08
Authors
Huang, Mian
Contributor
Advisor
Department
Chemistry
Instructor
Depositor
Speaker
Researcher
Consultant
Interviewer
Annotator
Journal Title
Journal ISSN
Volume Title
Publisher
Volume
Number/Issue
Starting Page
Ending Page
Alternative Title
Abstract
Recently scientists are interested in developing new variants of red fluorescent proteins (RFPs), because biosensors basing on RFPs show visual superiority in live-cell imaging. In this thesis, the structures of two RFP variants, K78BoF and mRuby0.4-3, were investigated by X-ray crystallography. A monomeric crystal structure of K78BoF was obtained at pH 6 with resolution at 2.13Å, while a dimeric crystal structure of mRuby0.4-3 was determined at pH 8.2 with resolution at 2.63Å. Both of their chromophores are formed by Met-Tyr-Gly with K78BoF in the cis state and mRuby0.4-3 in trans. Hydrogen bonds connecting water molecules and residues to K78BoF chromophore probably stabilize the conformation of the chromophore and contribute to the red fluorescence emitted. However, the substitution of an unnatural amino acid pboronophenylalanine breaks two hydrogen bonds, causing a deviation of the conformation. Analysis of hydrogen bonds on mRuby0.4-3 chromophore was limited by the relatively low resolution of the crystal.
Description
Keywords
Red fluorescent protein, Crystallization, X-ray diffraction, Chromophore
Citation
Extent
Format
Geographic Location
Time Period
Related To
Table of Contents
Rights
All UHM dissertations and theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
Rights Holder
Local Contexts
Email libraryada-l@lists.hawaii.edu if you need this content in ADA-compliant format.