English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Aromatic amino acids are critical for stability of the bicoid homeodomain

Subramaniam, V., Jovin, T. M., & Rivera-Pomar, R. (2001). Aromatic amino acids are critical for stability of the bicoid homeodomain. Journal of Biological Chemistry, 276: DOI 10.1074/jbc.M102292200, pp. 21506-21511.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F729-0 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-E0C9-4
Genre: Journal Article

Files

show Files
hide Files
:
600078.pdf (Publisher version), 174KB
Name:
600078.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Subramaniam, V.1, Author              
Jovin, T. M.1, Author              
Rivera-Pomar, R.2, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              
2Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              

Content

show
hide
Free keywords: Nuclear-magnetic-resonance; DNA-binding specificity; Ultraviolet circular-dichroism; Antennapedia homeodomain; Engrailed homeodomain; Angstrom resolution; Drosophila embryo; Recognition helix; Buried tryptophan; Crystal-structure
 Abstract: The Drosophila Bicoid (Bcd) protein plays a dual role as a transcription and translation factor dependent on the unique DNA and RNA binding properties of the homeodomain (HD). We have used circular dichroism and fluorescence spectroscopy to probe the structure and stability of the Bcd-HD, for which a high resolution structure is not yet available. The fluorescence from the single tryptophan residue in the HD (Trp-48) is strongly quenched in the native state but is dramatically enhanced (;20-fold) upon denaturation. Similar results were obtained with the Ultrabithorax HD (Ubx-HD), suggesting that the unusual tryptophan fluorescence may be a general phenomenon of HD proteins. We have used site-directed mutagenesis to explore the role of aromatic acids in the structure of the Bcd-HD and to evaluate the proposal that interactions between the strictly conserved Trp residue in HDs and nearby aromatic residues are responsible for the fluorescence quenching in the native state. We determined that both Trp-48 and Phe-8 in the N-terminal region of the HD are individually necessary for structural stability of the Bcd-HD, the latter most likely as a factor coordinating the orientation of the N-terminal helix I and the recognition helix for efficient binding to a DNA target.

Details

show
hide
Language(s): eng - English
 Dates: 2001
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 236989
Other: 34689
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of Biological Chemistry
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 276 Sequence Number: DOI 10.1074/jbc.M102292200 Start / End Page: 21506 - 21511 Identifier: -