Analyzing the weak dimerization of a cellulose binding module by analytical ultracentrifugation

Dmitrii Fedorov, Piotr Batys, David B. Hayes, Maria Sammalkorpi, Markus B. Linder*

*Tämän työn vastaava kirjoittaja

Tutkimustuotos: LehtiartikkeliArticleScientificvertaisarvioitu

4 Sitaatiot (Scopus)
80 Lataukset (Pure)

Abstrakti

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.

AlkuperäiskieliEnglanti
Sivut1995-2004
Sivumäärä10
JulkaisuInternational Journal of Biological Macromolecules
Vuosikerta163
DOI - pysyväislinkit
TilaJulkaistu - 15 marrask. 2020
OKM-julkaisutyyppiA1 Julkaistu artikkeli, soviteltu

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