PDBsum entry 4id4

Hydrolase

PDB id: 4id4

Contents

Protein chain

263 a.a.

Metals

_MG

_CL ×5

Waters ×425

PDB id:

4id4

Name:

Hydrolase

Title:

Crystal structure of chimeric beta-lactamase ctem-17m

Structure:

Beta-lactamase tem, beta-lactamase pse-4. Chain: a. Synonym: irt-4, penicillinase, tem-1, tem-16/caz-7, tem-2, tem- 24/caz-6, tem-3, tem-4, tem-5, tem-6, tem-8/caz-2, beta-lactamase carb-1, carbenicillinase 1. Engineered: yes

Source:

Escherichia coli, pseudomonas aeruginosa. Organism_taxid: 562, 287. Gene: bla, blat-3, blat-4, blat-5, blat-6, carb1, pse4. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.05Å R-factor: 0.116 R-free: 0.138

Authors:

J.Park,S.Gobeil,J.N.Pelletier,A.M.Berghuis

Key ref:

S.M.Gobeil et al. (2014). Maintenance of native-like protein dynamics may not be required for engineering functional proteins. Chem Biol, 21, 1330-1340. PubMed id: 25200606 DOI: 10.1016/j.chembiol.2014.07.016

Date:

11-Dec-12 Release date: 25-Dec-13

Protein chain

P16897  (BLP4_PSEAI) -  Beta-lactamase PSE-4 from Pseudomonas aeruginosa

Seq: 288 a.a.

Struc: 263 a.a.*

Protein chain

P62593  (BLAT_ECOLX) -  Beta-lactamase TEM from Escherichia coli

Seq: 286 a.a.

Struc: 263 a.a.*

* PDB and UniProt seqs differ at 150 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.5.2.6 - beta-lactamase.
• Pathway: Penicillin Biosynthesis and Metabolism
• Reaction: a beta-lactam + H2O = a substituted beta-amino acid
• Cofactor: Zn(2+)

DOI no: 10.1016/j.chembiol.2014.07.016

Chem Biol 21:1330-1340 (2014)

PubMed id: 25200606

Maintenance of native-like protein dynamics may not be required for engineering functional proteins.

S.M.Gobeil, C.M.Clouthier, J.Park, D.Gagné, A.M.Berghuis, N.Doucet, J.N.Pelletier.

ABSTRACT

Proteins are dynamic systems, and understanding dynamics is critical for fully understanding protein function. Therefore, the question of whether laboratory engineering has an impact on protein dynamics is of general interest. Here, we demonstrate that two homologous, naturally evolved enzymes with high degrees of structural and functional conservation also exhibit conserved dynamics. Their similar set of slow timescale dynamics is highly restricted, consistent with evolutionary conservation of a functionally important feature. However, we also show that dynamics of a laboratory-engineered chimeric enzyme obtained by recombination of the two homologs exhibits striking difference on the millisecond timescale, despite function and high-resolution crystal structure (1.05 Å) being conserved. The laboratory-engineered chimera is thus functionally tolerant to modified dynamics on the timescale of catalytic turnover. Tolerance to dynamic variation implies that maintenance of native-like protein dynamics may not be required when engineering functional proteins.