Loessl, Philip, Koelbel, Knut, Taenzler, Dirk, Nannemann, David ORCID: 0000-0002-4946-3263, Ihling, Christian H., Keller, Manuel V., Schneider, Marian, Zaucke, Frank, Meiler, Jens ORCID: 0000-0001-8945-193X and Sinz, Andrea (2014). Analysis of Nidogen-1/Laminin gamma 1 Interaction by Cross-Linking, Mass Spectrometry, and Computational Modeling Reveals Multiple Binding Modes. PLoS One, 9 (11). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203
Full text not available from this repository.Abstract
We describe the detailed structural investigation of nidogen-1/laminin gamma 1 complexes using full-length nidogen-1 and a number of laminin gamma 1 variants. The interactions of nidogen-1 with laminin variants gamma 1 LEb2-4, gamma 1 LEb2-4 N836D, gamma 1 short arm, and gamma 1 short arm N836D were investigated by applying a combination of (photo-) chemical cross-linking, high-resolution mass spectrometry, and computational modeling. In addition, surface plasmon resonance and ELISA studies were used to determine kinetic constants of the nidogen-1/laminin gamma 1 interaction. Two complementary cross-linking strategies were pursued to analyze solution structures of laminin gamma 1 variants and nidogen-1. The majority of distance information was obtained with the homobifunctional amine-reactive cross-linker bis(sulfosuccinimidyl) glutarate. In a second approach, UV-induced cross-linking was performed after incorporation of the diazirine-containing unnatural amino acids photo-leucine and photo-methionine into laminin gamma 1 LEb2-4, laminin gamma 1 short arm, and nidogen-1. Our results indicate that Asn-836 within laminin gamma 1 LEb3 domain is not essential for complex formation. Cross-links between laminin gamma 1 short arm and nidogen-1 were found in all protein regions, evidencing several additional contact regions apart from the known interaction site. Computational modeling based on the cross-linking constraints indicates the existence of a conformational ensemble of both the individual proteins and the nidogen-1/laminin gamma 1 complex. This finding implies different modes of interaction resulting in several distinct protein-protein interfaces.
Item Type: | Journal Article | ||||||||||||||||||||||||||||||||||||||||||||
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URN: | urn:nbn:de:hbz:38-423445 | ||||||||||||||||||||||||||||||||||||||||||||
DOI: | 10.1371/journal.pone.0112886 | ||||||||||||||||||||||||||||||||||||||||||||
Journal or Publication Title: | PLoS One | ||||||||||||||||||||||||||||||||||||||||||||
Volume: | 9 | ||||||||||||||||||||||||||||||||||||||||||||
Number: | 11 | ||||||||||||||||||||||||||||||||||||||||||||
Date: | 2014 | ||||||||||||||||||||||||||||||||||||||||||||
Publisher: | PUBLIC LIBRARY SCIENCE | ||||||||||||||||||||||||||||||||||||||||||||
Place of Publication: | SAN FRANCISCO | ||||||||||||||||||||||||||||||||||||||||||||
ISSN: | 1932-6203 | ||||||||||||||||||||||||||||||||||||||||||||
Language: | English | ||||||||||||||||||||||||||||||||||||||||||||
Faculty: | Unspecified | ||||||||||||||||||||||||||||||||||||||||||||
Divisions: | Unspecified | ||||||||||||||||||||||||||||||||||||||||||||
Subjects: | no entry | ||||||||||||||||||||||||||||||||||||||||||||
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URI: | http://kups.ub.uni-koeln.de/id/eprint/42344 |
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