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Title: Crystallization of the class IV adenylyl cyclase from Yersinia pestis

Abstract

The class IV adenylyl cyclase from Y. pestis has been crystallized in an orthorhombic form suitable for structure determination. The class IV adenylyl cyclase from Yersinia pestis has been cloned and crystallized in both a triclinic and an orthorhombic form. An amino-terminal His-tagged construct, from which the tag was removed by thrombin, crystallized in a triclinic form diffracting to 1.9 Å, with one dimer per asymmetric unit and unit-cell parameters a = 33.5, b = 35.5, c = 71.8 Å, α = 88.7, β = 82.5, γ = 65.5°. Several mutants of this construct crystallized but diffracted poorly. A non-His-tagged native construct (179 amino acids, MW = 20.5 kDa) was purified by conventional chromatography and crystallized in space group P2{sub 1}2{sub 1}2{sub 1}. These crystals have unit-cell parameters a = 56.8, b = 118.6, c = 144.5 Å, diffract to 3 Å and probably have two dimers per asymmetric unit and V{sub M} = 3.0 Å{sup 3} Da{sup −1}. Both crystal forms appear to require pH below 5, complicating attempts to incorporate nucleotide ligands into the structure. The native construct has been produced as a selenomethionine derivative and crystallized for phasing and structure determination.

Authors:
; ; ;  [1]
  1. Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8310 (United States)
Publication Date:
OSTI Identifier:
22356296
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 3; Other Information: PMCID: PMC2197185; PMID: 16511301; PUBLISHER-ID: ll5043; OAI: oai:pubmedcentral.nih.gov:2197185; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; DIMERS; LIGANDS; SPACE GROUPS

Citation Formats

Smith, Natasha, Kim, Sook-Kyung, Reddy, Prasad T., and Gallagher, D. Travis, E-mail: travis.gallagher@nist.gov. Crystallization of the class IV adenylyl cyclase from Yersinia pestis. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106002855.
Smith, Natasha, Kim, Sook-Kyung, Reddy, Prasad T., & Gallagher, D. Travis, E-mail: travis.gallagher@nist.gov. Crystallization of the class IV adenylyl cyclase from Yersinia pestis. United Kingdom. doi:10.1107/S1744309106002855.
Smith, Natasha, Kim, Sook-Kyung, Reddy, Prasad T., and Gallagher, D. Travis, E-mail: travis.gallagher@nist.gov. Wed . "Crystallization of the class IV adenylyl cyclase from Yersinia pestis". United Kingdom. doi:10.1107/S1744309106002855.
@article{osti_22356296,
title = {Crystallization of the class IV adenylyl cyclase from Yersinia pestis},
author = {Smith, Natasha and Kim, Sook-Kyung and Reddy, Prasad T. and Gallagher, D. Travis, E-mail: travis.gallagher@nist.gov},
abstractNote = {The class IV adenylyl cyclase from Y. pestis has been crystallized in an orthorhombic form suitable for structure determination. The class IV adenylyl cyclase from Yersinia pestis has been cloned and crystallized in both a triclinic and an orthorhombic form. An amino-terminal His-tagged construct, from which the tag was removed by thrombin, crystallized in a triclinic form diffracting to 1.9 Å, with one dimer per asymmetric unit and unit-cell parameters a = 33.5, b = 35.5, c = 71.8 Å, α = 88.7, β = 82.5, γ = 65.5°. Several mutants of this construct crystallized but diffracted poorly. A non-His-tagged native construct (179 amino acids, MW = 20.5 kDa) was purified by conventional chromatography and crystallized in space group P2{sub 1}2{sub 1}2{sub 1}. These crystals have unit-cell parameters a = 56.8, b = 118.6, c = 144.5 Å, diffract to 3 Å and probably have two dimers per asymmetric unit and V{sub M} = 3.0 Å{sup 3} Da{sup −1}. Both crystal forms appear to require pH below 5, complicating attempts to incorporate nucleotide ligands into the structure. The native construct has been produced as a selenomethionine derivative and crystallized for phasing and structure determination.},
doi = {10.1107/S1744309106002855},
journal = {Acta Crystallographica. Section F},
number = Pt 3,
volume = 62,
place = {United Kingdom},
year = {Wed Mar 01 00:00:00 EST 2006},
month = {Wed Mar 01 00:00:00 EST 2006}
}