PDBsum entry 4zwi

Lyase/lyase inhibitor

PDB id: 4zwi

Contents

Protein chain

257 a.a.

Ligands

DMS ×7

62I

Metals

_ZN

Waters ×271

PDB id:

4zwi

Name:

Lyase/lyase inhibitor

Title:

Surface lysine acetylated human carbonic anhydrase ii in complex with a sulfamate-based inhibitor

Structure:

Carbonic anhydrase 2. Chain: a. Synonym: carbonate dehydratase ii,carbonic anhydrasE C,cac,carbonic anhydrase ii,ca-ii. Engineered: yes. Other_details: acetylated lysine at position 112

Source:

Homo sapiens. Human. Organism_taxid: 9606. Cell_line: e. Coli. Gene: ca2. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.60Å R-factor: 0.181 R-free: 0.225

Authors:

C.L.Lomelino,B.P.Mahon,M.Mckenna

Key ref:

B.P.Mahon et al. (2015). Observed surface lysine acetylation of human carbonic anhydrase II expressed in Escherichia coli. Protein Sci, 24, 1800-1807. PubMed id: 26266677 DOI: 10.1002/pro.2771

Date:

19-May-15 Release date: 26-Aug-15

Protein chain

P00918  (CAH2_HUMAN) -  Carbonic anhydrase 2 from Homo sapiens

Seq: 260 a.a.

Struc: 257 a.a.*

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

Enzyme reactions

• Enzyme class 2: E.C.4.2.1.1 - carbonic anhydrase.
• Reaction: hydrogencarbonate + H⁺ = CO2 + H2O
• Cofactor: Zn(2+)
• Enzyme class 3: E.C.4.2.1.69 - cyanamide hydratase.
• Reaction: urea = cyanamide + H2O

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1002/pro.2771

Protein Sci 24:1800-1807 (2015)

PubMed id: 26266677

Observed surface lysine acetylation of human carbonic anhydrase II expressed in Escherichia coli.

B.P.Mahon, C.L.Lomelino, A.L.Salguero, J.M.Driscoll, M.A.Pinard, R.McKenna.

ABSTRACT

Acetylation of surface lysine residues of proteins has been observed in Escherichia coli (E. coli), an organism that has been extensively utilized for recombinant protein expression. This post-translational modification is shown to be important in various processes such as metabolism, stress-response, transcription, and translation. As such, utilization of E. coli expression systems for protein production may yield non-native acetylation events of surface lysine residues. Here we present the crystal structures of wild-type and a variant of human carbonic anhydrase II (hCA II) that have been expressed in E. coli and exhibit surface lysine acetylation and we speculate on the effect this has on the conformational stability of each enzyme. Both structures were determined to 1.6 Å resolution and show clear electron density for lysine acetylation. The lysine acetylation does not distort the structure and the surface lysine acetylation events most likely do not interfere with the biological interpretation. However, there is a reduction in conformational stability in the hCA II variant compared to wild type (∼4°C decrease). This may be due to other lysine acetylation events that have occurred but are not visible in the crystal structure due to intrinsic disorder. Therefore, surface lysine acetylation events may affect overall protein stability and crystallization, and should be considered when using E. coli expression systems.