Mapping an Equilibrium Folding Intermediate of the Cytolytic Pore Toxin ClyA with Single-Molecule FRET

The Journal of Physical Chemistry. B
Fabian DingfelderBenjamin Schuler

Abstract

The 303-residue cytolytic toxin ClyA forms a stable α-helical monomer. In the presence of detergents or membranes, however, the protein makes a large conformational transition to the protomer state, which is competent for assembly into a dodecameric cytolytic pore. In this study, we map the structure of the ClyA monomer during denaturant-induced unfolding with single-molecule Förster resonance energy transfer (FRET) spectroscopy. To this end, we probe intramolecular distances of six different segments of ClyA by placing donor and acceptor fluorophores at corresponding positions along the chain. We identify an intermediate state that contains the folded core consisting of three of the α-helices that make up the helical bundle present in the structure of both the monomer and the protomer, but with the C- and N-terminal helices unfolded, in accord with the secondary structure content estimated from circular dichroism (CD) spectroscopy. The existence of this intermediate is likely to be a consequence of the structural bistability underlying the biological function of ClyA: The terminal helices are part of the largest rearrangements during protomer formation, and the local differences in stability we detect may prime the protein for...Continue Reading

References

Jan 1, 1979·Methods in Enzymology·A HeleniusC Tanford
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·Y GotoA L Fink
Jan 1, 1987·Progress in Biophysics and Molecular Biology·R Jaenicke
Jan 1, 1995·Annual Review of Biophysics and Biomolecular Structure·A L Fink
Aug 15, 2000·Annual Review of Biophysics and Biomolecular Structure·W A EatonJ Hofrichter
Mar 4, 2003·Proceedings of the National Academy of Sciences of the United States of America·Elizabeth RhoadesGilad Haran
Jan 1, 1997·Annual Review of Physical Chemistry·T PlakhotnikU P Wild
Apr 23, 2004·Current Opinion in Structural Biology·José Nelson Onuchic, Peter G Wolynes
Aug 23, 2005·Biophysical Journal·Barbara K MüllerDon C Lamb
Jan 13, 2006·The Journal of Physical Chemistry. a·Maksim KouzaD Thirumalai
May 11, 2006·Chemical Reviews·Xavier MichaletMarcus Jäger
May 11, 2006·Chemical Reviews·Valerie Daggett
Jun 7, 2006·The Journal of Biological Chemistry·Svetomir B TzokovPer A Bullough
Sep 8, 2006·Methods in Molecular Biology·Benjamin Schuler
Feb 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Daniel NettelsBenjamin Schuler
Apr 5, 2007·Nature Protocols·Norma J Greenfield
Oct 16, 2007·Biophysical Journal·Zdenek Petrásek, Petra Schwille
Jan 29, 2008·Current Opinion in Structural Biology·Benjamin Schuler, William A Eaton
Mar 28, 2008·Proceedings of the National Academy of Sciences of the United States of America·Robbyn L TuinstraBrian F Volkman
Apr 17, 2008·Annual Review of Biochemistry·Alessandro BorgiaJane Clarke
Jun 10, 2009·Proceedings of the National Academy of Sciences of the United States of America·Yann GambinAshok A Deniz
Sep 11, 2009·The Journal of Chemical Physics·Irina V GopichAttila Szabo
Jan 12, 2010·The Journal of Chemical Physics·Carlo Camilloni, Ludovico Sutto
Mar 3, 2010·Annual Review of Biophysics·D ThirumalaiChangbong Hyeon
May 6, 2010·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Anastasia LomanJörg Enderlein
Apr 8, 2011·Bioinformatics·Corinna VehlowMichael Lappe
Oct 13, 2011·Nature Communications·Menahem PirchiGilad Haran
May 3, 2012·Proceedings of the National Academy of Sciences of the United States of America·Irina V Gopich, Attila Szabo
Sep 18, 2012·Proceedings of the National Academy of Sciences of the United States of America·Hagen HofmannBenjamin Schuler
Nov 13, 2012·Nature Methods·Stanislav KalininClaus A M Seidel
Nov 28, 2012·Science·Ken A Dill, Justin L MacCallum
Jan 2, 2013·Methods in Enzymology·Christoph PieperJörg Enderlein
Jan 3, 2013·Current Opinion in Structural Biology·Gabriel Zoldák, Matthias Rief
Jan 15, 2013·Current Opinion in Structural Biology·Benjamin Schuler, Hagen Hofmann
Aug 8, 2013·Journal of the American Chemical Society·Misha SoskineGiovanni Maglia
Dec 18, 2013·Chemical Society Reviews·Priya R Banerjee, Ashok A Deniz
Jan 18, 2014·Chemical Reviews·Marco BrucaleBruno Samorì
Feb 6, 2015·Nature Communications·Stephan BenkeBenjamin Schuler

❮ Previous
Next ❯

Citations

Nov 12, 2020·Structure·Allen K Kim, Lauren L Porter
Jul 20, 2019·Current Opinion in Structural Biology·Georg KrainerMichael Schlierf
Sep 2, 2021·JACS Au·Fabian DingfelderBenjamin Schuler
Oct 26, 2021·Biopolymers·Andy LiWangLee-Ping Wang
Nov 25, 2021·Journal of the American Chemical Society·Samuel Naudi-FabraSigrid Milles

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.