6V3T

X-Ray Crystal Structure of Tribolium castaneum Arylalkylamine N-acyltransferase in Complex with Acetyl-CoA

Summary for 6V3T

• Entry DOI: 10.2210/pdb6v3t/pdb
• Descriptor: Dopamine N-acetyltransferase-like Protein, ACETYL COENZYME *A (3 entities in total)
• Functional Keywords: aanat, complex, coa, transferase
• Biological source: Tribolium castaneum (Red flour beetle)
• Total number of polymer chains: 6
• Total formula weight: 153166.70

Authors

Lewandowski, E.M.,Chen, Y. (deposition date: 2019-11-26, release date: 2020-02-05, Last modification date: 2024-11-06)

Primary citation

O'Flynn, B.G.,Lewandowski, E.M.,Prins, K.C.,Suarez, G.,McCaskey, A.N.,Rios-Guzman, N.M.,Anderson, R.L.,Shepherd, B.A.,Gelis, I.,Leahy, J.W.,Chen, Y.,Merkler, D.J.
Characterization of ArylalkylamineN-Acyltransferase fromTribolium castaneum: An Investigation into a Potential Next-Generation Insecticide Target.
Acs Chem.Biol., 15:513-523, 2020

PubMed Abstract: The growing issue of insecticide resistance has meant the identification of novel insecticide targets has never been more important. Arylalkylamine -acyltransferases (AANATs) have been suggested as a potential new target. These promiscuous enzymes are involved in the -acylation of biogenic amines to form -acylamides. In insects, this process is a key step in melanism, hardening of the cuticle, removal of biogenic amines, and in the biosynthesis of fatty acid amides. The unique nature of each AANAT isoform characterized indicates each organism accommodates an assembly of discrete AANATs relatively exclusive to that organism. This implies a high potential for selectivity in insecticide design, while also maintaining polypharmacology. Presented here is a thorough kinetic and structural analysis of AANAT found in one of the most common secondary pests of all plant commodities in the world, The enzyme, named AANAT0, catalyzes the formation of short-chain -acylarylalkylamines, with short-chain acyl-CoAs (C2-C10), benzoyl-CoA, and succinyl-CoA functioning in the role of acyl donor. Recombinant AANAT0 was expressed and purified from and was used to investigate the kinetic and chemical mechanism of catalysis. The kinetic mechanism is an ordered sequential mechanism with the acyl-CoA binding first. pH-rate profiles and site-directed mutagenesis studies identified amino acids critical to catalysis, providing insights about the chemical mechanism of AANAT0. A crystal structure was obtained for AANAT0 bound to acetyl-CoA, revealing valuable information about its active site. This combination of kinetic analysis and crystallography alongside mutagenesis and sequence analysis shines light on some approaches possible for targeting AANAT0 and other AANATs for novel insecticide design.

PubMed: 31967772
DOI: 10.1021/acschembio.9b00973

Experimental method

X-RAY DIFFRACTION (2.84 Å)