PDBsum entry 5amx

Hydrolase

PDB id: 5amx

Contents

Protein chain

279 a.a.

Ligands

SO4

Waters ×359

PDB id:

5amx

Name:

Hydrolase

Title:

Crystal structure of proteinase k processed with the crystaldirect automated mounting and cryo-cooling technology

Structure:

Proteinase k. Chain: a. Fragment: unp residues 106-384. Synonym: endopeptidase k, tritirachium alkaline proteinase. Ec: 3.4.21.64

Source:

Engyodontium album. Organism_taxid: 37998

Resolution:

1.01Å R-factor: 0.158 R-free: 0.166

Authors:

U.Zander,G.Hoffmann,I.Cornaciu,F.Cipriani,J.A.Marquez

Key ref:

U.Zander et al. (2016). Automated harvesting and processing of protein crystals through laser photoablation. Acta Crystallogr D Struct Biol, 72, 454-466. PubMed id: 27050125 DOI: 10.1107/S2059798316000954

Date:

02-Sep-15 Release date: 13-Apr-16

Protein chain

P06873  (PRTK_PARAQ) -  Proteinase K from Parengyodontium album

Seq: 384 a.a.

Struc: 279 a.a.

Enzyme reactions

• Enzyme class: E.C.3.4.21.64 - peptidase K.
• Reaction: Hydrolysis of keratin and of other proteins, with subtilisin-like specificity. Hydrolyzes peptides amides.

DOI no: 10.1107/S2059798316000954

Acta Crystallogr D Struct Biol 72:454-466 (2016)

PubMed id: 27050125

Automated harvesting and processing of protein crystals through laser photoablation.

U.Zander, G.Hoffmann, I.Cornaciu, J.P.Marquette, G.Papp, C.Landret, G.Seroul, J.Sinoir, M.Röwer, F.Felisaz, S.Rodriguez-Puente, V.Mariaule, P.Murphy, M.Mathieu, F.Cipriani, J.A.Márquez.

ABSTRACT

Currently, macromolecular crystallography projects often require the use of highly automated facilities for crystallization and X-ray data collection. However, crystal harvesting and processing largely depend on manual operations. Here, a series of new methods are presented based on the use of a low X-ray-background film as a crystallization support and a photoablation laser that enable the automation of major operations required for the preparation of crystals for X-ray diffraction experiments. In this approach, the controlled removal of the mother liquor before crystal mounting simplifies the cryocooling process, in many cases eliminating the use of cryoprotectant agents, while crystal-soaking experiments are performed through diffusion, precluding the need for repeated sample-recovery and transfer operations. Moreover, the high-precision laser enables new mounting strategies that are not accessible through other methods. This approach bridges an important gap in automation and can contribute to expanding the capabilities of modern macromolecular crystallography facilities.