This old version of Proteopedia is provided for student assignments while the new version is undergoing repairs. Content and edits done in this old version of Proteopedia after March 1, 2026 will eventually be lost when it is retired in about June of 2026.

Apply for new accounts at the new Proteopedia. Your logins will work in both the old and new versions.


1xrk

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
Line 1: Line 1:
-
[[Image:1xrk.gif|left|200px]]
+
{{Seed}}
 +
[[Image:1xrk.png|left|200px]]
<!--
<!--
Line 9: Line 10:
{{STRUCTURE_1xrk| PDB=1xrk | SCENE= }}
{{STRUCTURE_1xrk| PDB=1xrk | SCENE= }}
-
'''Crystal structure of a mutant bleomycin binding protein from Streptoalloteichus hindustanus displaying increased thermostability'''
+
===Crystal structure of a mutant bleomycin binding protein from Streptoalloteichus hindustanus displaying increased thermostability===
-
==Overview==
+
<!--
-
Limited thermostability of antibiotic resistance markers has restricted genetic research in the field of extremely thermophilic Archaea and bacteria. In this study, we used directed evolution and selection in the thermophilic bacterium Thermus thermophilus HB27 to find thermostable variants of a bleomycin-binding protein from the mesophilic bacterium Streptoalloteichus hindustanus. In a single selection round, we identified eight clones bearing five types of double mutated genes that provided T. thermophilus transformants with bleomycin resistance at 77 degrees C, while the wild-type gene could only do so up to 65 degrees C. Only six different amino acid positions were altered, three of which were glycine residues. All variant proteins were produced in Escherichia coli and analyzed biochemically for thermal stability and functionality at high temperature. A synthetic mutant resistance gene with low GC content was designed that combined four substitutions. The encoded protein showed up to 17 degrees C increased thermostability and unfolded at 85 degrees C in the absence of bleomycin, whereas in its presence the protein unfolded at 100 degrees C. Despite these highly thermophilic properties, this mutant was still able to function normally at mesophilic temperatures in vivo. The mutant protein was co-crystallized with bleomycin, and the structure of the binary complex was determined to a resolution of 1.5 A. Detailed structural analysis revealed possible molecular mechanisms of thermostabilization and enhanced antibiotic binding, which included the introduction of an intersubunit hydrogen bond network, improved hydrophobic packing of surface indentations, reduction of loop flexibility, and alpha-helix stabilization. The potential applicability of the thermostable selection marker is discussed.
+
The line below this paragraph, {{ABSTRACT_PUBMED_15640151}}, adds the Publication Abstract to the page
 +
(as it appears on PubMed at http://www.pubmed.gov), where 15640151 is the PubMed ID number.
 +
-->
 +
{{ABSTRACT_PUBMED_15640151}}
==About this Structure==
==About this Structure==
Line 31: Line 35:
[[Category: Ligand binding protein]]
[[Category: Ligand binding protein]]
[[Category: Thermostable mutant]]
[[Category: Thermostable mutant]]
-
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 15:25:03 2008''
+
 
 +
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 14:39:45 2008''

Revision as of 11:39, 29 July 2008

Image:1xrk.png

Template:STRUCTURE 1xrk

Crystal structure of a mutant bleomycin binding protein from Streptoalloteichus hindustanus displaying increased thermostability

Template:ABSTRACT PUBMED 15640151

About this Structure

1XRK is a Single protein structure of sequence from Streptoalloteichus hindustanus. Full crystallographic information is available from OCA.

Reference

Engineering a selectable marker for hyperthermophiles., Brouns SJ, Wu H, Akerboom J, Turnbull AP, de Vos WM, van der Oost J, J Biol Chem. 2005 Mar 25;280(12):11422-31. Epub 2005 Jan 7. PMID:15640151

Page seeded by OCA on Tue Jul 29 14:39:45 2008

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/1xrk"
Personal tools