PDBsum entry 6n4u

Hydrolase

PDB id: 6n4u

Contents

Protein chain

279 a.a.

Ligands

SO4

Metals

_CA ×2

Waters ×23

PDB id:

6n4u

Name:

Hydrolase

Title:

Microed structure of proteinase k at 2.75a resolution from a single milled crystal.

Structure:

Proteinase k. Chain: a. Synonym: endopeptidase k,tritirachium alkaline proteinase. Engineered: yes

Source:

Engyodontium album. Organism_taxid: 37998. Gene: prok. Expressed in: engyodontium album. Expression_system_taxid: 37998

Authors:

M.W.Martynowycz,W.Zhao,J.Hattne,G.J.Jensen,T.Gonen

Key ref:

M.W.Martynowycz et al. (2019). Collection of Continuous Rotation MicroED Data from Ion Beam-Milled Crystals of Any Size. Structure, 27, 545. PubMed id: 30661853 DOI: 10.1016/j.str.2018.12.003

Date:

20-Nov-18 Release date: 06-Feb-19

Protein chain

P06873  (PRTK_PARAQ) -  Proteinase K from Parengyodontium album

Seq: 384 a.a.

Struc: 279 a.a.*

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

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: 10.1016/j.str.2018.12.003

Structure 27:545 (2019)

PubMed id: 30661853

Collection of Continuous Rotation MicroED Data from Ion Beam-Milled Crystals of Any Size.

M.W.Martynowycz, W.Zhao, J.Hattne, G.J.Jensen, T.Gonen.

ABSTRACT

Microcrystal electron diffraction (MicroED) allows for macromolecular structure solution from nanocrystals. To create crystals of suitable size for MicroED data collection, sample preparation typically involves sonication or pipetting a slurry of crystals from a crystallization drop. The resultant crystal fragments are fragile and the quality of the data that can be obtained from them is sensitive to subsequent sample preparation for cryoelectron microscopy as interactions in the water-air interface can damage crystals during blotting. Here, we demonstrate the use of a focused ion beam to generate lamellae of macromolecular protein crystals for continuous rotation MicroED that are of ideal thickness, easy to locate, and require no blotting optimization. In this manner, crystals of nearly any size may be scooped and milled to desired dimensions prior to data collection, thus streamlining the methodology for sample preparation for MicroED.