Document Type
Article
Language
eng
Format of Original
13 p.
Publication Date
10-2016
Publisher
Wiley
Source Publication
Protein Science
Source ISSN
0961-8368
Abstract
Urea amidolyase (UAL) is a multifunctional biotin-dependent enzyme that contributes to both bacterial and fungal pathogenicity by catalyzing the ATP-dependent cleavage of urea into ammonia and CO2. UAL is comprised of two enzymatic components: urea carboxylase (UC) and allophanate hydrolase (AH). These enzyme activities are encoded on separate but proximally related genes in prokaryotes while, in most fungi, they are encoded by a single gene that produces a fusion enzyme on a single polypeptide chain. It is unclear whether the UC and AH activities are connected through substrate channeling or other forms of direct communication. Here, we use multiple biochemical approaches to demonstrate that there is no substrate channeling or interdomain/intersubunit communication between UC and AH. Neither stable nor transient interactions can be detected between prokaryotic UC and AH and the catalytic efficiencies of UC and AH are independent of one another. Furthermore, an artificial fusion of UC and AH does not significantly alter the AH enzyme activity or catalytic efficiency. These results support the surprising functional independence of AH from UC in both the prokaryotic and fungal UAL enzymes and serve as an important reminder that the evolution of multifunctional enzymes through gene fusion events does not always correlate with enhanced catalytic function.
Recommended Citation
Lin, Yi; Boese, Cody J.; and St. Maurice, Martin, "The Urea Carboxylase and Allophanate Hydrolase Activities of Urea Amidolyase Are Functionally Independent" (2016). Biological Sciences Faculty Research and Publications. 535.
https://epublications.marquette.edu/bio_fac/535
Comments
Accepted version. Protein Science, Vol. 25, No. 10 (October 2016): 1812-1824. DOI. © 2016 John Wiley & Sons, Inc. Used with permission.