Purification, crystallization, and preliminary X-ray diffraction analysis of the Tricorn protease hexamer from Thermoplasma acidophilum

Jürgen Bosch, Tomohiro Tamura, Gleb Bourenkov, Wolfgang Baumeister, Lars Oliver Essen

Research output: Contribution to journalArticle

Abstract

Tricorn protease from Thermoplasma acidophilum is a hexameric enzyme; in vivo the hexamers assemble further to form large icosahedral capsids of 14.6 MDa. Recombinant Tricorn protease was purified as an enzymatically active hexamer of 0.72 MDa that formed crystals of octahedral morphology under low-ionic-strength conditions. These crystals belong to space group C2 with unit cell dimensions a = 307.5 Å, b = 163.2 Å, c = 220.9 Å, β = 105.5° and diffract to 2.2-Å resolution using high-brilliance synchrotron radiation. Based on analysis of the self-rotation function and the presence of a pseudo-origin peak in the native Patterson map, a packing model was derived for the complex, comprising 1.5 hexamers per asymmetric unit with a solvent content of 43%. Due to the ninefold noncrystallographic symmetry the Tricorn crystals represent an interesting case for phasing X-ray crystallographic data by electron microscopic phase information.

Original languageEnglish (US)
Pages (from-to)83-87
Number of pages5
JournalJournal of Structural Biology
Volume134
Issue number1
DOIs
StatePublished - 2001
Externally publishedYes

Keywords

  • Electron microscopy
  • Self-compartmentalizing protease
  • Thermophile
  • Tricorn
  • X-ray crystallography

ASJC Scopus subject areas

  • Structural Biology

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