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          Institute: MPI für Biochemie     Collection: Structural Biology (W. Baumeister)     Display Documents

ID: 34916.0, MPI für Biochemie / Structural Biology (W. Baumeister)
Specific orientation and two-dimensional crystallization of the proteasome at metal-chelating lipid interfaces
Authors:Thess, A.; Hutschenreiter, S.; Hofmann, M.; Tampe, R.; Baumeister, W.; Guckenberger, R.
Date of Publication (YYYY-MM-DD):2002-09-27
Title of Journal:Journal of Biological Chemistry
Journal Abbrev.:J. Biol. Chem.
Issue / Number:39
Start Page:36321
End Page:36328
Review Status:Peer-review
Audience:Not Specified
Abstract / Description: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.
Free Keywords:20S
Comment of the Author/Creator:Date: 2002, SEP 27
6.2.2007: 20S added to keywords
External Publication Status:published
Document Type:Article
Communicated by:Anton Hillebrand
Affiliations:MPI für Biochemie/Structural Biology (W. Baumeister)/Research Group Guckenberger
MPI für Biochemie/Structural Biology (W. Baumeister)
External Affiliations:Univ Frankfurt, Biozentrum, Inst Biochem, D-60439 Frankfurt, Germany
Identifiers:ISI:000178275100069 [ID No:1]
ISSN:0021-9258 [ID No:2]
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