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  Specific orientation and two-dimensional crystallization of the proteasome at metal-chelating lipid interfaces

Thess, A., Hutschenreiter, S., Hofmann, M., Tampe, R., Baumeister, W., & Guckenberger, R. (2002). Specific orientation and two-dimensional crystallization of the proteasome at metal-chelating lipid interfaces. Journal of Biological Chemistry, 277(39), 36321-36328.

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Genre: Journal Article
Alternative Title : J. Biol. Chem.

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 Creators:
Thess, A., Author
Hutschenreiter, S., Author
Hofmann, M., Author
Tampe, R., Author
Baumeister, W.1, Author              
Guckenberger, R.1, Author              
Affiliations:
1Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

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Free keywords: 20S
 Abstract: The potential of a protein-engineered His tag to immobilize macromolecules in a predictable orientation at metal-chelating lipid interfaces was investigated using recombinant 20 S proteasomes His-tagged in various positions. Electron micrographs demonstrated that the orientation of proteasomes bound to chelating lipid films could be controlled via the location of their His tags: proteasomes His-tagged at their sides displayed exclusively side-on views, while proteasomes His-tagged at their ends displayed exclusively end-on views. The activity of proteasomes immobilized at chelating lipid interfaces was well preserved. In solution, His-tagged proteasomes hydrolyzed casein at rates comparable with wild- type proteasomes, unless the His tags were located in the vicinity of the N termini of alpha-subunits. The N termini of a-subunits might partly occlude the entrance channel in a-rings through which substrates enter the proteasome for subsequent degradation. A combination of electron micrographs and atomic force microscope topographs revealed a propensity of vertically oriented proteasomes to crystallize in two dimensions on fluid lipid films. The oriented immobilization of His-tagged proteins at biocompatible lipid interfaces will assist structural studies as well as the investigation of biomolecular interaction via a wide variety of surface-sensitive techniques including single-molecule analysis.

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Language(s): eng - English
 Dates: 2002-09-27
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 34916
ISI: 000178275100069
 Degree: -

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Title: Journal of Biological Chemistry
  Alternative Title : J. Biol. Chem.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 277 (39) Sequence Number: - Start / End Page: 36321 - 36328 Identifier: ISSN: 0021-9258