Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 19933-19942 |
| Number of pages | 10 |
| Journal | Nanoscale |
| Volume | 11 |
| Issue number | 42 |
| DOIs | |
| Publication status | Published - 14 Nov 2019 |
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Conformation switching of single native proteins revealed by nanomechanical probing without a pulling force. / Gutierrez Mejia, F.A.; Moerland, Christian; van Ijzendoorn, Leo; Prins, Menno (Corresponding author).
In: Nanoscale, Vol. 11, No. 42, 14.11.2019, p. 19933-19942.Research output: Contribution to journal › Article › Academic › peer-review
TY - JOUR
T1 - Conformation switching of single native proteins revealed by nanomechanical probing without a pulling force
AU - Gutierrez Mejia, F.A.
AU - Moerland, Christian
AU - van Ijzendoorn, Leo
AU - Prins, Menno
PY - 2019/11/14
Y1 - 2019/11/14
N2 - Protein conformational changes are essential to biological function, and the heterogeneous nature of the corresponding protein states provokes an interest to measure conformational changes at the single molecule level. Here we demonstrate that conformational changes in single native proteins can be revealed by non-covalent antibody-targeting of specific domains within the protein, using nanomechanical probing without an applied pulling force. The protein of interest was captured between a particle and a substrate and three properties were quantified: the twist amplitude related to an applied torque, torsional compliance related to rotational Brownian motion, and translational Brownian displacement. Calcium-dependent conformation switching was studied in native human cardiac troponin, a heterotrimer protein complex that regulates the contraction and relaxation of heart muscle cells and is also a key biomarker for diagnosing myocardial infarction. The data reveal a change in mechanical properties upon conformation switching from the non-saturated to the calcium-saturated state, which in cardiomyocytes gives myosin motor proteins access to actin filaments. A clear increase was observed in the molecular stiffness for the calcium-saturated protein conformation. Using libraries of monoclonal antibodies, the nanomechanical probing of conformation by antibody targeting opens avenues for characterizing single native protein complexes for research as well as for diagnostic applications.
AB - Protein conformational changes are essential to biological function, and the heterogeneous nature of the corresponding protein states provokes an interest to measure conformational changes at the single molecule level. Here we demonstrate that conformational changes in single native proteins can be revealed by non-covalent antibody-targeting of specific domains within the protein, using nanomechanical probing without an applied pulling force. The protein of interest was captured between a particle and a substrate and three properties were quantified: the twist amplitude related to an applied torque, torsional compliance related to rotational Brownian motion, and translational Brownian displacement. Calcium-dependent conformation switching was studied in native human cardiac troponin, a heterotrimer protein complex that regulates the contraction and relaxation of heart muscle cells and is also a key biomarker for diagnosing myocardial infarction. The data reveal a change in mechanical properties upon conformation switching from the non-saturated to the calcium-saturated state, which in cardiomyocytes gives myosin motor proteins access to actin filaments. A clear increase was observed in the molecular stiffness for the calcium-saturated protein conformation. Using libraries of monoclonal antibodies, the nanomechanical probing of conformation by antibody targeting opens avenues for characterizing single native protein complexes for research as well as for diagnostic applications.
UR - http://www.scopus.com/inward/record.url?scp=85074379992&partnerID=8YFLogxK
U2 - 10.1039/C9NR01448A
DO - 10.1039/C9NR01448A
M3 - Article
VL - 11
SP - 19933
EP - 19942
JO - Nanoscale
JF - Nanoscale
SN - 2040-3364
IS - 42
ER -