PDBsum entry 4lcm

Transferase

PDB id: 4lcm

Contents

Protein chains

393 a.a.

Waters ×12

PDB id:

4lcm

Name:

Transferase

Title:

Simvastatin synthase (lovd), from aspergillus terreus, lovd9 mutant (simh9014)

Structure:

Transesterase. Chain: a, b, c, d. Engineered: yes. Mutation: yes

Source:

Aspergillus terreus. Organism_taxid: 33178. Gene: lovd. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

3.19Å R-factor: 0.210 R-free: 0.252

Authors:

X.Gao,M.R.Sawaya,T.O.Yeates,Y.Tang

Key ref:

G.Jiménez-Osés et al. (2014). The role of distant mutations and allosteric regulation on LovD active site dynamics. Nat Chem Biol, 10, 431-436. PubMed id: 24727900 DOI: 10.1038/nchembio.1503

Date:

21-Jun-13 Release date: 02-Apr-14

Protein chains

Q9Y7D1  (LOVD_ASPTE) -  Monacolin J acid methylbutanoyltransferase from Aspergillus terreus

Seq: 413 a.a.

Struc: 393 a.a.*

* PDB and UniProt seqs differ at 28 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.3.1.238 - monacolin J acid methylbutanoate transferase.
• Reaction: monacolin J carboxylate + (S)-2-methylbutanoyl-[2-methylbutanoate polyketide synthase] = lovastatin carboxylate + holo-[2-methylbutanoate polyketide synthase]

DOI no: 10.1038/nchembio.1503

Nat Chem Biol 10:431-436 (2014)

PubMed id: 24727900

The role of distant mutations and allosteric regulation on LovD active site dynamics.

G.Jiménez-Osés, S.Osuna, X.Gao, M.R.Sawaya, L.Gilson, S.J.Collier, G.W.Huisman, T.O.Yeates, Y.Tang, K.N.Houk.

ABSTRACT

Natural enzymes have evolved to perform their cellular functions under complex selective pressures, which often require their catalytic activities to be regulated by other proteins. We contrasted a natural enzyme, LovD, which acts on a protein-bound (LovF) acyl substrate, with a laboratory-generated variant that was transformed by directed evolution to accept instead a small free acyl thioester and no longer requires the acyl carrier protein. The resulting 29-mutant variant is 1,000-fold more efficient in the synthesis of the drug simvastatin than the wild-type LovD. This is to our knowledge the first nonpatent report of the enzyme currently used for the manufacture of simvastatin as well as the intermediate evolved variants. Crystal structures and microsecond-scale molecular dynamics simulations revealed the mechanism by which the laboratory-generated mutations free LovD from dependence on protein-protein interactions. Mutations markedly altered conformational dynamics of the catalytic residues, obviating the need for allosteric modulation by the acyl carrier LovF.