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In Vitro Approaches for the Assessment of Serpin Polymerization

Elliston, ELK; Lomas, DA; Irving, JA; (2018) In Vitro Approaches for the Assessment of Serpin Polymerization. Methods in Molecular Biology , 1826 pp. 87-107. 10.1007/978-1-4939-8645-3_6. Green open access

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Abstract

Serpin polymerization is the result of end-to-end ordered aggregation of serpin monomers into linear unbranched chains. This change in molecular state represents the basis of several conformational diseases with pathological gain-of-function and loss-of-function phenotypes, termed serpinopathies. Tools that enable quantification and characterization of polymer formation are therefore important to the study of serpin behavior in this pathophysiological context. Such methods rely on different manifestations of molecular change: polymerization-the generation of molecules with increasing molecular weight-is accompanied by concomitant structural rearrangements in the constituent subunits. Different approaches may be appropriate dependent on whether measurements are made on static samples, such as tissue or cell culture extracts, or in time-resolved experiments, often undertaken using polymers artificially induced under in vitro destabilizing conditions. In the former category, we describe the application of polyacrylamide electrophoresis, Western blot, ELISA, and negative-stain electron microscopy and in the latter category, Förster resonance energy transfer and fluorescence spectroscopy using environment-sensitive probes.

Type: Article
Title: In Vitro Approaches for the Assessment of Serpin Polymerization
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1007/978-1-4939-8645-3_6
Publisher version: http://dx.doi.org/10.1007/978-1-4939-8645-3_6
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: ELISA, FRET, Monoclonal antibody, Negative-stain electron microscopy, Oligomerization, Polymerization, Protein aggregation, Western blot
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Respiratory Medicine
UCL > Provost and Vice Provost Offices > VP: Health
URI: https://discovery.ucl.ac.uk/id/eprint/10059668
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