dc.contributor.author |
Nevzorova T. |
|
dc.contributor.author |
Zhao Q. |
|
dc.contributor.author |
Lomakin Y. |
|
dc.contributor.author |
Ponomareva A. |
|
dc.contributor.author |
Mukhitov A. |
|
dc.contributor.author |
Purohit P. |
|
dc.contributor.author |
Weisel J. |
|
dc.contributor.author |
Litvinov R. |
|
dc.date.accessioned |
2018-09-19T23:12:57Z |
|
dc.date.available |
2018-09-19T23:12:57Z |
|
dc.date.issued |
2017 |
|
dc.identifier.issn |
2191-1630 |
|
dc.identifier.uri |
https://dspace.kpfu.ru/xmlui/handle/net/146081 |
|
dc.description.abstract |
© 2016, Springer Science+Business Media New York.Interactions of DNA with proteins are essential for key biological processes and have both a fundamental and practical significance. In particular, DNA binding to anti-DNA antibodies is a pathogenic mechanism in autoimmune pathology, such as systemic lupus erythematosus. Here we measured at the single-molecule level binding and forced unbinding of surface-attached DNA and a monoclonal anti-DNA antibody MRL4 from a lupus erythematosus mouse. In optical trap-based force spectroscopy, a microscopic antibody-coated latex bead is trapped by a focused laser beam and repeatedly brought into contact with a DNA-coated surface. After careful discrimination of non-specific interactions, we showed that the DNA-antibody rupture force spectra had two regimes, reflecting formation of weaker (20–40 pN) and stronger (>40 pN) immune complexes that implies the existence of at least two bound states with different mechanical stability. The two-dimensional force-free off-rate for the DNA-antibody complexes was ∼2.2 × 10−3 s−1, the transition state distance was ∼0.94 nm, the apparent on-rate was ∼5.26 s−1, and the stiffness of the DNA-antibody complex was characterized by a spring constant of 0.0021 pN/nm, suggesting that the DNA-antibody complex is a relatively stable, but soft and deformable macromolecular structure. The stretching elasticity of the DNA molecules was characteristic of single-stranded DNA, suggesting preferential binding of the MRL4 antibody to one strand of DNA. Collectively, the results provide fundamental characteristics of formation and forced dissociation of DNA-antibody complexes that help to understand principles of DNA-protein interactions and shed light on the molecular basis of autoimmune diseases accompanied by formation of anti-DNA antibodies. |
|
dc.relation.ispartofseries |
BioNanoScience |
|
dc.subject |
Anti-DNA antibody |
|
dc.subject |
DNA |
|
dc.subject |
Nanomechanics |
|
dc.subject |
Optical trap |
|
dc.subject |
Single-molecule force spectroscopy |
|
dc.subject |
Two-dimensional kinetics |
|
dc.title |
Single-Molecule Interactions of a Monoclonal Anti-DNA Antibody with DNA |
|
dc.type |
Article |
|
dc.relation.ispartofseries-issue |
1 |
|
dc.relation.ispartofseries-volume |
7 |
|
dc.collection |
Публикации сотрудников КФУ |
|
dc.relation.startpage |
132 |
|
dc.source.id |
SCOPUS21911630-2017-7-1-SID85017510298 |
|