Document Detail


Toward detecting and identifying macromolecules in a cellular context: template matching applied to electron tomograms.
MedLine Citation:
PMID:  11087814     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Electron tomography is the only technique available that allows us to visualize the three-dimensional structure of unfixed and unstained cells currently with a resolution of 6-8 nm, but with the prospect to reach 2-4 nm. This raises the possibility of detecting and identifying specific macromolecular complexes within their cellular context by virtue of their structural signature. Templates derived from the high-resolution structure of the molecule under scrutiny are used to search the reconstructed volume. Here we outline and test a computationally feasible two-step procedure: In a first step, mean-curvature motion is used for segmentation, yielding subvolumes that contain with a high probability macromolecules in the expected size range. Subsequently, the particles contained in the subvolumes are identified by cross-correlation, using a set of three-dimensional templates. With simulated and real tomographic data we demonstrate that such an approach is feasible and we explore the detection limits. Even structurally similar particles, such as the thermosome, GroEL, and the 20S proteasome can be identified with high fidelity. This opens up exciting prospects for mapping the territorial distribution of macromolecules and for analyzing molecular interactions in situ.
Authors:
J Bohm; A S Frangakis; R Hegerl; S Nickell; D Typke; W Baumeister
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  97     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2000 Dec 
Date Detail:
Created Date:  2001-01-16     Completed Date:  2001-02-15     Revised Date:  2010-09-14    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  14245-50     Citation Subset:  IM    
Affiliation:
Department of Molecular Structural Biology, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18a, D-82152 Martinsried, Germany. boehm@biochem.mpg.de
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MeSH Terms
Descriptor/Qualifier:
Algorithms
Chaperonin 60 / chemistry*,  isolation & purification
Cysteine Endopeptidases / chemistry*,  isolation & purification
Macromolecular Substances
Models, Molecular
Multienzyme Complexes / chemistry*,  isolation & purification
Proteasome Endopeptidase Complex
Protein Conformation
Tomography, X-Ray Computed / methods
Chemical
Reg. No./Substance:
0/Chaperonin 60; 0/Macromolecular Substances; 0/Multienzyme Complexes; EC 3.4.22.-/Cysteine Endopeptidases; EC 3.4.25.1/Proteasome Endopeptidase Complex
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