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Cellulose binding modules (CBMs) are found widely in different proteins that act on cellulose. Because they allow a very easy way of binding recombinant proteins to cellulose, they have become widespread in many biotechnological applications involving cellulose. One commonly used variant is the CBMCipA from Clostridium thermocellum. Here we studied the oligomerization behavior of CBMCipA, as such solution association may have an impact on its use. As the principal approach, we used sedimentation velocity and sedimentation equilibrium analytical ultracentrifugation. To enhance our understanding of the possible interactions, we used molecular dynamics simulations. By analysis of the sedimentation velocity data by a discrete model genetic algorithm and by building a binding isotherm based on weight average sedimentation coefficient and by global fitting of sedimentation equilibrium data we found that the CBMCipA shows a weak dimerization interaction with a dissociation constant KD of 90 ± 30 μM. As the KD of CBMCipA binding to cellulose is below 1 μM, we conclude that the dimerization is unlikely to affect cellulose binding. However, at high concentrations used in some applications of the CBMCipA, its dimerization is likely to have a marked effect on its solution behavior.
|Number of pages||10|
|Journal||International Journal of Biological Macromolecules|
|Publication status||Published - 15 Nov 2020|
|MoE publication type||A1 Journal article-refereed|
- Analytical ultracentrifugation
- Protein engineering
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the Academy of Finland's Centre of Excellence in Molecular Engineering of Biosynthetic Hybrid Materials research (2014-2019), HYBER
01/01/2017 → 31/12/2019
Project: Academy of Finland: Other research funding