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Balancing specificity and promiscuity in enzyme evolution: multidimensional activity transitions in the alkaline phosphatase superfamily

DOI: 10.1021/jacs.8b10290 DOI Help

Authors: Bert Van Loo (University of Cambridge) , Christopher D. Bayer (University of Cambridge) , Gerhard Fischer (University of Cambridge) , Stefanie Jonas (University of Cambridge) , Eugene Valkov (University of Cambridge) , Mark F. Mohamed (University of Cambridge) , Anastassia Vorobieva (University of Cambridge) , Celine Dutruel (University of Cambridge) , Marko Hyvonen (University of Cambridge) , Florian Hollfelder (University of Cambridge)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of The American Chemical Society

State: Published (Approved)
Published: November 2018
Diamond Proposal Number(s): 6889

Abstract: Highly proficient, promiscuous enzymes can be springboards for functional evolution, able to avoid loss of function during adaptation by their capacity to promote multiple reactions. We employ systematic comparative study of structure, sequence and substrate specificity to track the evolution of specificity and reactivity between promiscuous members of clades of the alkaline phosphatase (AP) superfamily. Construction of a phylogenetic tree of protein sequences maps out the likely transition zone between arylsulfatases (ASs) and phosphonate monoester hydrolases (PMHs). Kinetic analysis shows that all enzymes characterized have four chemically distinct phospho- and sulfoesterase activities, with rate accelerations ranging from 1011-1017-fold for their primary and 109-1012-fold for their promiscuous reactions, suggesting that catalytic promiscuity is widespread in the AP-superfamily. This functional characterization and crystallography reveal a novel class of ASs that is so similar in sequence to known PMHs that it had not been recognized as having diverged in function. Based on analysis of snapshots of catalytic promiscuity ‘in transition’ we develop possible models that would allow functional evolution and determine scenarios for trade-off between multiple activities. For the new ASs we observe largely invariant substrate specificity that would facilitate the transition from ASs to PMHs via trade-off-free molecular exaptation, i.e. evolution without initial loss of primary activity and specificity toward the original substrate. This ability to bypass low activity generalists provides a molecular solution to avoid adaptive conflict.

Journal Keywords: Anions; Peptides and proteins; Monomers; Organophosphorus compounds; Chemical specificity

Diamond Keywords: Enzymes

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography

Added On: 03/12/2018 11:19

Discipline Tags:

Catalysis Life Sciences & Biotech Structural biology Chemistry Biochemistry

Technical Tags:

Diffraction Macromolecular Crystallography (MX)