title: Evolution of enzymatic activity in the tautomerase superfamily: mechanistic and structural studies of the 1,3-dichloropropene catabolic enzymes.
abstract: The use of the soil fumigant Telone II, which contains a mixture of cis- and trans-1,3-dichloropropene, to control plant-parasitic nematodes is a common agricultural practice for maximizing yields of various crops. The effectiveness of Telone II is limited by the rapid turnover of the dichloropropenes in the soil due to the presence of bacterial catabolic pathways, which may be of recent origin. The characterization of three enzymes in these pathways, trans-3-chloroacrylic acid dehalogenase (CaaD), cis-3-chloroacrylic acid dehalogenase (cis-CaaD), and malonate semialdehyde decarboxylase (MSAD), has uncovered intriguing catalytic mechanisms as well as a fascinating evolutionary lineage for these proteins. Sequence comparisons and mutagenesis studies revealed that all three enzymes belong to the tautomerase superfamily. Tautomerase superfamily members with known structures are characterized by a beta-alpha-beta structural fold. Moreover, they have a conserved N-terminal proline, which plays an important catalytic role. Mechanistic, NMR, and pH rate studies of the two dehalogenases, coupled with a crystal structure of CaaD inactivated by 3-bromopropiolate, indicate that they use a general acid/base mechanism to catalyze the conversion of their respective isomer of 3-chloroacrylate to malonate semialdehyde. The reaction is initiated by the conjugate addition of water to the C-2, C-3 double bond and is followed by the loss of HCl. MSAD processes malonate semialdehyde to acetaldehyde, and is the first identified decarboxylase in the tautomerase superfamily. The catalytic mechanism is not well defined but the N-terminal proline plays a prominent role and may function as a general acid catalyst, similar to its role in CaaD and cis-CaaD. These are the first structural and mechanistic details for tautomerase superfamily members that catalyze either a hydration or a decarboxylation reaction, rather than a tautomerization reaction, in which Pro-1 serves as a general acid catalyst rather than as a general base catalyst. The available information on the 1,3-dichloropropene catabolic enzymes allows speculation on the possible evolutionary origins of their activities.
authors.affiliation: Division of Medicinal Chemistry, College of Pharmacy, The University of Texas, Austin, TX 78712-1074, USA.
authors.forename: Gerrit J
authors.initials: GJ
authors.lastname: Poelarends
authors.forename: Christian P
authors.initials: CP
authors.lastname: Whitman
mesh_terms: D000498:Allyl Compounds / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites / Q000502:physiology; D001673:Biodegradation, Environmental; D004789:Enzyme Activation; D019143:Evolution, Molecular; D006843:Hydrocarbons, Chlorinated; D007535:Isomerases / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D010105:Oxygenases / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011549:Pseudomonas / Q000201:enzymology
chemical_list: D000498:Allyl Compounds; D006843:Hydrocarbons, Chlorinated; C023891:1,3-dichloro-1-propene; D010105:Oxygenases; D007535:Isomerases