9 research outputs found
Safe distances of separation and space-planning safety measures in the prevention of the effects of accidents in 'Seveso' plants
10.00; Translated from German (CEE-SEVESO Workshop, Luxembourg, 16-18 Nov 1988)SIGLEAvailable from British Library Document Supply Centre- DSC:9022.381(HSE-Trans--13063)T / BLDSC - British Library Document Supply CentreGBUnited Kingdo
Automated Assignment of MS/MS Cleavable Cross-Links in Protein 3D-Structure Analysis
CID-MS/MS cleavable cross-linkers hold an enormous potential for an automated analysis of cross-linked products, which is essential for conducting structural proteomics studies. The created characteristic fragment ion patterns can easily be used for an automated assignment and discrimination of cross-linked products. To date, there are only a few software solutions available that make use of these properties, but none allows for an automated analysis of cleavable cross-linked products. The MeroX software fills this gap and presents a powerful tool for protein 3D-structure analysis in combination with MS/MS cleavable cross-linkers. We show that MeroX allows an automatic screening of characteristic fragment ions, considering static and variable peptide modifications, and effectively scores different types of cross-links. No manual input is required for a correct assignment of cross-links and false discovery rates are calculated. The self-explanatory graphical user interface of MeroX provides easy access for an automated cross-link search platform that is compatible with commonly used data file formats, enabling analysis of data originating from different instruments. The combination of an MS/MS cleavable cross-linker with a dedicated software tool for data analysis provides an automated workflow for 3D-structure analysis of proteins. MeroX is available at www.StavroX.com
Structural Insights into Retinal Guanylylcyclase–GCAP-2 Interaction Determined by Cross-Linking and Mass Spectrometry
The retinal guanylylcyclases ROS-GC 1 and 2 are regulated
via the intracellular site by guanylylcyclase-activating proteins
(GCAPs). The mechanisms of how GCAPs activate their target proteins
remain elusive as exclusively structures of nonactivating calcium-bound
GCAP-1 and -2 are available. In this work, we apply a combination
of chemical cross-linking with amine-reactive cross-linkers and photoaffinity
labeling followed by a mass spectrometric analysis of the created
cross-linked products to study the interaction between N-terminally
myristoylated GCAP-2 and a peptide derived from the catalytic domain
of full-length ROS-GC 1. In our studies, only a few cross-linked products
were obtained for calcium-bound GCAP-2, pointing to a well-defined
structure of the GCAP-2–GC peptide complex. A much larger number
of cross-links were detected in the absence of calcium, indicating
a high flexibility of calcium-free GCAP-2 in the complex with the
GC peptide. On the basis of the distance constraints imposed by the
cross-links, we were able to create a structural model of the calcium-loaded
complex between myristoylated GCAP-2 and the GC peptide