Document Type

Restricted

Advisor

Marc Zimmer

Publication Date

2019

Comments

This paper is restricted to Connecticut College on-campus users until 5/14/2020.

Abstract

Green fluorescent protein, initially cloned and expressed from the bioluminescent jellyfish, Aequorea victoria (avGFP), has a wide range of uses in cellular biology, one of which includes uses as a biological marker1. Variants of GFP exist, but some residues are highly conserved and necessary for appropriate chromophore formation. Some of these conserved residues are the glycine residues at positions 31, 33 and 35, though it they are not part of the tripeptide that forms the chromophore2 .

The objective of this honors study and 3 related independent studies was to use computational methods to find out why the glycines at positions 31, 33, and 35 are so highly conserved in all fluorescent proteins and what role they play in chromophore formation, folding and stabilizing the protein. Precyclized immature structures (i.e. with no chromophore) have been used in our simulations because there is been evidence that conserved glycine residues play an important role in protein folding3 or chromophore formation, which occurs prior to cyclization. Although mutating the glycine at position 35 to a cysteine3 has been found to be somewhat fluorescent, we chose to make a less aggressive mutations, G31A, G33A, and G35A in order to study because alanine is the most structurally similar to glycine.

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The views expressed in this paper are solely those of the author.