PDBsum entry 4jm5

Oxidoreductase

PDB id: 4jm5

Contents

Protein chain

289 a.a.

Ligands

HEM

25T-MES

Waters ×516

PDB id:

4jm5

Name:

Oxidoreductase

Title:

Crystal structure of cytochromE C peroxidase w191g-gateless in complex with 2-amino-5-methylthiazole

Structure:

CytochromE C peroxidase. Chain: a. Fragment: residues 72-362, deletions g192-a193. Engineered: yes. Mutation: yes

Source:

Saccharomyces cerevisiae. Baker's yeast. Organism_taxid: 285006. Strain: rm11-1a. Gene: ccp1 ccp cpo ykr066c, scrg_04081. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

1.26Å R-factor: 0.129 R-free: 0.155

Authors:

S.E.Boyce,M.Fischer,I.Fish,B.K.Shoichet

Key ref:

G.J.Rocklin et al. (2013). Blind prediction of charged ligand binding affinities in a model binding site. J Mol Biol, 425, 4569-4583. PubMed id: 23896298 DOI: 10.1016/j.jmb.2013.07.030

Date:

13-Mar-13 Release date: 01-May-13

Protein chain

B3LRE1  (B3LRE1_YEAS1) -  Peroxidase from Saccharomyces cerevisiae (strain RM11-1a)

Seq: 362 a.a.

Struc: 289 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.1.11.1.- - ?????

DOI no: 10.1016/j.jmb.2013.07.030

J Mol Biol 425:4569-4583 (2013)

PubMed id: 23896298

Blind prediction of charged ligand binding affinities in a model binding site.

G.J.Rocklin, S.E.Boyce, M.Fischer, I.Fish, D.L.Mobley, B.K.Shoichet, K.A.Dill.

ABSTRACT

Predicting absolute protein-ligand binding affinities remains a frontier challenge in ligand discovery and design. This becomes more difficult when ionic interactions are involved because of the large opposing solvation and electrostatic attraction energies. In a blind test, we examined whether alchemical free-energy calculations could predict binding affinities of 14 charged and 5 neutral compounds previously untested as ligands for a cavity binding site in cytochrome c peroxidase. In this simplified site, polar and cationic ligands compete with solvent to interact with a buried aspartate. Predictions were tested by calorimetry, spectroscopy, and crystallography. Of the 15 compounds predicted to bind, 13 were experimentally confirmed, while 4 compounds were false negative predictions. Predictions had a root-mean-square error of 1.95kcal/mol to the experimental affinities, and predicted poses had an average RMSD of 1.7Å to the crystallographic poses. This test serves as a benchmark for these thermodynamically rigorous calculations at predicting binding affinities for charged compounds and gives insights into the existing sources of error, which are primarily electrostatic interactions inside proteins. Our experiments also provide a useful set of ionic binding affinities in a simplified system for testing new affinity prediction methods.