Abstract
The CH3 homodimer at the C-terminal end of the antibody heavy chain is the key noncovalent interaction stabilizing antibody proteins. Here, we use single-molecule force spectroscopy to investigate the dissociation mechanics of CH3 as a proxy for antibody mechanical stability. We find the CH3 homodimer to be a highly stable complex, and its dissociation force of >150 pN at a loading rate of ≈5500 pN/s exceeds the stability of most protein-protein interactions studied to date. Separated C H3 monomers, on the other hand, are mechanically labile and only short-lived. Each CH3 monomer contains a conserved buried disulfide bridge, and we find that the successive reduction of one or both disulfide bridges in the dimer results in a stepwise decrease of the dissociation force. This suggests a structural role of the disulfide bridges helping to mold the high-affinity domain-domain interface, even though they are neither required for nor directly involved in dimerization. Taken together, our results set a limit on how much force a single antibody can bear and reveal the CH3 homodimer as a mechanical fastener that prevents antibody dissociation.
| Original language | English |
|---|---|
| Pages (from-to) | 15085-15091 |
| Number of pages | 7 |
| Journal | Journal of the American Chemical Society |
| Volume | 135 |
| Issue number | 40 |
| DOIs | |
| Publication status | Published - 2013 |
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ASJC Scopus subject areas
- Chemistry(all)
- Catalysis
- Biochemistry
- Colloid and Surface Chemistry
Cite this
Mechanical stability of the antibody domain CH3 homodimer in different oxidation states. / Bertz, Morten; Buchner, Johannes; Rief, Matthias.
In: Journal of the American Chemical Society, Vol. 135, No. 40, 2013, p. 15085-15091.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mechanical stability of the antibody domain CH3 homodimer in different oxidation states
AU - Bertz, Morten
AU - Buchner, Johannes
AU - Rief, Matthias
PY - 2013
Y1 - 2013
N2 - The CH3 homodimer at the C-terminal end of the antibody heavy chain is the key noncovalent interaction stabilizing antibody proteins. Here, we use single-molecule force spectroscopy to investigate the dissociation mechanics of CH3 as a proxy for antibody mechanical stability. We find the CH3 homodimer to be a highly stable complex, and its dissociation force of >150 pN at a loading rate of ≈5500 pN/s exceeds the stability of most protein-protein interactions studied to date. Separated C H3 monomers, on the other hand, are mechanically labile and only short-lived. Each CH3 monomer contains a conserved buried disulfide bridge, and we find that the successive reduction of one or both disulfide bridges in the dimer results in a stepwise decrease of the dissociation force. This suggests a structural role of the disulfide bridges helping to mold the high-affinity domain-domain interface, even though they are neither required for nor directly involved in dimerization. Taken together, our results set a limit on how much force a single antibody can bear and reveal the CH3 homodimer as a mechanical fastener that prevents antibody dissociation.
AB - The CH3 homodimer at the C-terminal end of the antibody heavy chain is the key noncovalent interaction stabilizing antibody proteins. Here, we use single-molecule force spectroscopy to investigate the dissociation mechanics of CH3 as a proxy for antibody mechanical stability. We find the CH3 homodimer to be a highly stable complex, and its dissociation force of >150 pN at a loading rate of ≈5500 pN/s exceeds the stability of most protein-protein interactions studied to date. Separated C H3 monomers, on the other hand, are mechanically labile and only short-lived. Each CH3 monomer contains a conserved buried disulfide bridge, and we find that the successive reduction of one or both disulfide bridges in the dimer results in a stepwise decrease of the dissociation force. This suggests a structural role of the disulfide bridges helping to mold the high-affinity domain-domain interface, even though they are neither required for nor directly involved in dimerization. Taken together, our results set a limit on how much force a single antibody can bear and reveal the CH3 homodimer as a mechanical fastener that prevents antibody dissociation.
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U2 - 10.1021/ja405076j
DO - 10.1021/ja405076j
M3 - Article
VL - 135
SP - 15085
EP - 15091
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 40
ER -