PDBsum entry 3ef3

Hydrolase

PDB id: 3ef3

Contents

Protein chain

196 a.a. *

Ligands

NXC

Waters ×322

* Residue conservation analysis

PDB id:

3ef3

Name:

Hydrolase

Title:

Cut-1a; ncn-pt-pincer-cutinase hybrid

Structure:

Cutinase-1. Chain: a. Synonym: cutin hydrolase 1. Engineered: yes. Mutation: yes

Source:

Fusarium solani f. Sp. Pisi. Organism_taxid: 70791. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.50Å R-factor: 0.160 R-free: 0.178

Authors:

L.Rutten,J.P.B.A.Mannie,M.Lutz,P.Gros

Key ref:

L.Rutten et al. (2009). Solid-state structural characterization of cutinase-ECE-pincer-metal hybrids. Chemistry, 15, 4270-4280. PubMed id: 19219875

Date:

08-Sep-08 Release date: 28-Jul-09

Protein chain

P00590  (CUTI1_FUSVN) -  Cutinase 1 from Fusarium vanettenii

Seq: 230 a.a.

Struc: 196 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.1.1.74 - cutinase.
• Reaction: cutin + H2O = cutin monomers

Chemistry 15:4270-4280 (2009)

PubMed id: 19219875

Solid-state structural characterization of cutinase-ECE-pincer-metal hybrids.

L.Rutten, B.Wieczorek, J.P.Mannie, C.A.Kruithof, H.P.Dijkstra, M.R.Egmond, M.Lutz, R.J.Klein Gebbink, P.Gros, G.van Koten.

ABSTRACT

The first crystal structures of lipases that have been covalently modified through site-selective inhibition by different organometallic phosphonate-pincer-metal complexes are described. Two ECE-pincer-type d(8)-metal complexes, that is, platinum (1) or palladium (2) with phosphonate esters (ECE = [(EtO)-(O=)P(-O-C(6)H(4)-(NO(2))-4)(-C(3)H(6)-4-(C(6)H(2)-(CH(2)E)(2))](-); E = NMe(2) or SMe) were introduced prior to crystallization and have been shown to bind selectively to the Ser(120) residue in the active site of the lipase cutinase to give cut-1 (platinum) or cut-2 (palladium) hybrids. For all five presented crystal structures, the ECE-pincer-platinum or -palladium head group sticks out of the cutinase molecule and is exposed to the solvent. Depending on the nature of the ECE-pincer-metal head group, the ECE-pincer-platinum and -palladium guests occupy different pockets in the active site of cutinase, with concomitant different stereochemistries on the phosphorous atom for the cut-1 (S(P)) and cut-2 (R(P)) structures. When cut-1 was crystallized under halide-poor conditions, a novel metal-induced dimeric structure was formed between two cutinase-bound pincer-platinum head groups, which are interconnected through a single mu-Cl bridge. This halide-bridged metal dimer shows that coordination chemistry is possible with protein-modified pincer-metal complexes. Furthermore, we could use NCN-pincer-platinum complex 1 as site-selective tool for the phasing of raw protein diffraction data, which shows the potential use of pincer-platinum complex 1 as a heavy-atom derivative in protein crystallography.