PDBsum entry: 1boh

Transferase

PDB id: 1boh

Contents

Protein chain

293 a.a. *

Waters ×63

* Residue conservation analysis

PDB id:

1boh

Name:

Transferase

Title:

Sulfur-substituted rhodanese (orthorhombic form)

Structure:

Rhodanese. Chain: a. Engineered: yes. Other_details: sulfur substituted at residue cys 247

Source:

Bos taurus. Cattle. Organism_taxid: 9913. Organ: liver. Cellular_location: cytoplasm. Expressed in: escherichia coli bl21. Expression_system_taxid: 511693.

Resolution:

2.30Å R-factor: 0.166 R-free: 0.215

Authors:

F.Gliubich,R.Berni,M.Cianci,R.J.Trevino,P.M.Horowitz, G.Zanotti

Key ref:

R.J.Trevino et al. (1999). NH2-terminal sequence truncation decreases the stability of bovine rhodanese, minimally perturbs its crystal structure, and enhances interaction with GroEL under native conditions. J Biol Chem, 274, 13938-13947. PubMed id: 10318804 DOI: 10.1074/jbc.274.20.13938

Date:

04-Aug-98 Release date: 27-Apr-99

Protein chain

P00586  (THTR_BOVIN) -  Thiosulfate sulfurtransferase

Seq: 297 a.a.

Struc: 293 a.a.

Enzyme reactions

• Enzyme class: E.C.2.8.1.1 - Thiosulfate sulfurtransferase.
• Reaction: Thiosulfate + cyanide = sulfite + thiocyanate

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

 Gene Ontology (GO) functional annotation 

• Cellular component: plasma membrane (4 terms)
• Biological process: rRNA transport (1 term)
• Biochemical function: transferase activity (4 terms)

reference

DOI no: 10.1074/jbc.274.20.13938

J Biol Chem 274:13938-13947 (1999)

PubMed id: 10318804

NH2-terminal sequence truncation decreases the stability of bovine rhodanese, minimally perturbs its crystal structure, and enhances interaction with GroEL under native conditions.

R.J.Trevino, F.Gliubich, R.Berni, M.Cianci, J.M.Chirgwin, G.Zanotti, P.M.Horowitz.

ABSTRACT

The NH2-terminal sequence of rhodanese influences many of its properties, ranging from mitochondrial import to folding. Rhodanese truncated by >9 residues is degraded in Escherichia coli. Mutant enzymes with lesser truncations are recoverable and active, but they show altered active site reactivities (Trevino, R. J., Tsalkova, T., Dramer, G., Hardesty, B., Chirgwin, J. M., and Horowitz, P. M. (1998) J. Biol. Chem. 273, 27841-27847), suggesting that the NH2-terminal sequence stabilizes the overall structure. We tested aspects of the conformations of these shortened species. Intrinsic and probe fluorescence showed that truncation decreased stability and increased hydrophobic exposure, while near UV CD suggested altered tertiary structure. Under native conditions, truncated rhodanese bound to GroEL and was released and reactivated by adding ATP and GroES, suggesting equilibrium between native and non-native conformers. Furthermore, GroEL assisted folding of denatured mutants to the same extent as wild type, although at a reduced rate. X-ray crystallography showed that Delta1-7 crystallized isomorphously with wild type in polyethyleneglycol, and the structure was highly conserved. Thus, the missing NH2-terminal residues that contribute to global stability of the native structure in solution do not significantly alter contacts at the atomic level of the crystallized protein. The two-domain structure of rhodanese was not significantly altered by drastically different crystallization conditions or crystal packing suggesting rigidity of the native rhodanese domains and the stabilization of the interdomain interactions by the crystal environment. The results support a model in which loss of interactions near the rhodanese NH2 terminus does not distort the folded native structure but does facilitate the transition in solution to a molten globule state, which among other things, can interact with molecular chaperones.

Selected figure(s)

Figure 3.
Fig. 3. Schematic drawing of the folding of rhodanese. Corresponding elements of secondary structure in domains I and II are denoted by capital letters. The sulfur atoms of the persulfide group at the active site are shown as black spheres. The drawing was obtained using the program MOLSCRIPT.

Figure 6.
Fig. 6. View of the cation-binding site in the crystal of wild type rhodanese, showing the distorted bipyramidal coordination of the cation. Atom symbols as in Fig. 5 and Table V. A very similar situation is present in the crystal of 1-7 mutant.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (1999, 274, 13938-13947) copyright 1999.

Figures were selected by an automated process.