title: A General Strategy for the Discovery of Metabolic Pathways: d-Threitol, l-Threitol, and Erythritol Utilization in Mycobacterium smegmatis.
abstract: We describe a general integrated bioinformatic and experimental strategy to discover the in vitro enzymatic activities and in vivo functions (metabolic pathways) of uncharacterized enzymes discovered in microbial genome projects using the ligand specificities of the solute binding proteins (SBPs) for ABC transporters. Using differential scanning fluorimetry, we determined that the SBP for an ABC transporter encoded by the genome of Mycobacterium smegmatis is stabilized by d-threitol. Using sequence similarity networks and genome neighborhood networks to guide selection of target proteins for pathway enzymes, we applied both in vitro and in vivo experimental approaches to discover novel pathways for catabolism of d-threitol, l-threitol, and erythritol.
authors.forename: Hua
authors.initials: H
authors.lastname: Huang
authors.forename: Michael S
authors.initials: MS
authors.lastname: Carter
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine , Bronx, New York 10461, United States.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Vetting
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine , Bronx, New York 10461, United States.
authors.forename: Nawar
authors.initials: N
authors.lastname: Al-Obaidi
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine , Bronx, New York 10461, United States.
authors.forename: Yury
authors.initials: Y
authors.lastname: Patskovsky
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine , Bronx, New York 10461, United States.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Almo
authors.forename: John A
authors.initials: JA
authors.lastname: Gerlt
mesh_terms: D004896:Erythritol / Q000378:metabolism*; D020102:Mycobacterium smegmatis / Q000378:metabolism*; D013237:Stereoisomerism; D013402:Sugar Alcohols / Q000378:metabolism*
chemical_list: D013402:Sugar Alcohols; C003460:threitol; D004896:Erythritol