PDBsum entry: 2rhh

Cell cycle

PDB id: 2rhh

Contents

Protein chain

304 a.a. *

Ligands

SO4

Waters ×114

* Residue conservation analysis

PDB id:

2rhh

Name:

Cell cycle

Title:

Synthetic gene encoded bacillus subtilis ftsz with bound sulfate ion

Structure:

Cell division protein ftsz. Chain: a. Fragment: residues 12-315. Engineered: yes

Source:

Bacillus subtilis. Organism_taxid: 1423. Gene: ftsz. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.00Å R-factor: 0.215 R-free: 0.257

Authors:

S.Lovell,Z.Halloran,K.Hjerrild,D.Sheridan,A.Burgin, L.Stewart,Accelerated Technologies Center For Gene To 3d Structure (Atcg3d)

Key ref:

A.Raymond et al. (2009). Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer. Bmc Biotechnol, 9, 37. PubMed id: 19383143 DOI: 10.1186/1472-6750-9-37

Date:

09-Oct-07 Release date: 21-Oct-08

Protein chain

P17865  (FTSZ_BACSU) -  Cell division protein FtsZ

Seq: 382 a.a.

Struc: 304 a.a.

 Gene Ontology (GO) functional annotation 

• Cellular component: protein complex (2 terms)
• Biological process: protein polymerization (1 term)
• Biochemical function: GTP binding (2 terms)

DOI no: 10.1186/1472-6750-9-37

Bmc Biotechnol 9:37 (2009)

PubMed id: 19383143

Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer.

A.Raymond, S.Lovell, D.Lorimer, J.Walchli, M.Mixon, E.Wallace, K.Thompkins, K.Archer, A.Burgin, L.Stewart.

ABSTRACT

BACKGROUND: With the goal of improving yield and success rates of heterologous protein production for structural studies we have developed the database and algorithm software package Gene Composer. This freely available electronic tool facilitates the information-rich design of protein constructs and their engineered synthetic gene sequences, as detailed in the accompanying manuscript. RESULTS: In this report, we compare heterologous protein expression levels from native sequences to that of codon engineered synthetic gene constructs designed by Gene Composer. A test set of proteins including a human kinase (P38alpha), viral polymerase (HCV NS5B), and bacterial structural protein (FtsZ) were expressed in both E. coli and a cell-free wheat germ translation system. We also compare the protein expression levels in E. coli for a set of 11 different proteins with greatly varied G:C content and codon bias. CONCLUSION: The results consistently demonstrate that protein yields from codon engineered Gene Composer designs are as good as or better than those achieved from the synonymous native genes. Moreover, structure guided N- and C-terminal deletion constructs designed with the aid of Gene Composer can lead to greater success in gene to structure work as exemplified by the X-ray crystallographic structure determination of FtsZ from Bacillus subtilis. These results validate the Gene Composer algorithms, and suggest that using a combination of synthetic gene and protein construct engineering tools can improve the economics of gene to structure research.