Authors

Benjamin Rajan

Type

Text

Type

Thesis

Advisor

Glynn, Steven. | French, Jarrod B

Date

2014-12-01

Keywords

Biochemistry | adenylosuccinate lyase, purinosome

Department

Department of Biochemistry and Cell Biology.

Language

en_US

Source

This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.

Identifier

http://hdl.handle.net/11401/76927

Publisher

The Graduate School, Stony Brook University: Stony Brook, NY.

Format

application/pdf

Abstract

Cells require purines for critical biological processes that can be created by the de novo biosynthetic pathway or maintained by the salvage pathways. The de novo pathway in humans requires 6 enzymes, some of which are multifunctional. Adenylosuccinate lyase (ASL) catalyzes step 8 of the de novo pathway converting susuccinylaminoimidazolecarboxamide ribonucleotide (SAICAR) to aminoimidazolecarboxamide ribotide (AICAR). It also catalyzes the second step of the purine nucleotide cycle by converting SAMP to AMP. A short isoform of ASL (sASL) is ubiquitously expressed in all cell types at a lower concentration. This shortened splice variant does not perform catalysis on either of the aforementioned substrates. It has been suggested that sASL may play an important structural role in the formation and stabilization of the purinosome assembly. The purpose of this work is to characterize the short isoform of ASL and elucidate its structure and function. To date, I have cloned sASL, expressed and purified the protein, and identified a buffer that stabilizes it. | 48 pages

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