4B52: Crystal Structure Of Gentlyase, The Neutral Metalloprotease Of Paenibacillus Polymyxa

Gentlyase is a bacterial extracellular metalloprotease that is widely applied in cell culture and for tissue dissociation and that belongs to the family of thermolysin-like proteases. The structure of thermolysin has been known since 1972 and that of Bacillus cereus neutral protease since 1992. However, the structure determination of other Bacillus neutral proteases has been hindered by their tendency to cannibalistic autolysis. High calcium conditions that allow the concentration and crystallization of the active Gentlyase metalloprotease without autoproteolysis were identified using thermal fluorescent shift assays. X-ray structures of the protease were solved in the absence and in the presence of the inhibitor phosphoramidon at 1.59 and 1.76 A resolution, respectively. No domain movement was observed upon inhibitor binding, although such movement is thought to be a general feature of the thermolysin-like protease family. Further analysis of the structure shows that the observed calcium dependency of Gentlyase stability may arise from a partly degenerated calcium site Ca1-2 and a deletion near site Ca3.
PDB ID: 4B52Download
MMDB ID: 106505
PDB Deposition Date: 2012/8/2
Updated in MMDB: 2013/03
Experimental Method:
x-ray diffraction
Resolution: 1.76  Å
Source Organism:
Similar Structures:
Biological Unit for 4B52: monomeric; determined by author and by software (PISA)
Molecular Components in 4B52
Label Count Molecule
Protein (1 molecule)
Molecule annotation
Chemicals (7 molecules)
* Click molecule labels to explore molecular sequence information.

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