A Rigid Hinge Region Is Necessary for High-Affinity Binding of Dimannose to Cyanovirin and Associated Constructs

Zhen Li, Ashini Bolia, Jason D. Maxwell, Andrey A. Bobkov, Giovanna Ghirlanda, S. Banu Ozkan, Claudio J. Margulis

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Mutations in the hinge region of cyanovirin-N (CVN) dictate its preferential oligomerization state. Constructs with the Pro51Gly mutation preferentially exist as monomers, whereas wild-type cyanovirin can form domain-swapped dimers under certain conditions. Because the hinge region is an integral part of the high-affinity binding site of CVN, we investigated whether this mutation affects the shape, flexibility, and binding affinity of domain B for dimannose. Our studies indicate that the capability of monomeric wild-type CVN to resist mechanical perturbations is enhanced when compared to that of constructs in which the hinge region is more flexible. Our computational results also show that enhanced flexibility leads to blocking of the binding site by allowing different rotational isomeric states of Asn53. Moreover, at higher temperatures, this observed flexibility leads to an interaction between Asn53 and Asn42, further hindering access to the binding site. On the basis of these results, we predicted that binding affinity for dimannose would be more favorable for cyanovirin constructs containing a wild-type hinge region, whereas affinity would be impaired in the case of mutants containing Pro51Gly. Experimental characterization by isothermal titration calorimetry of a set of cyanovirin mutants confirms this hypothesis. Those possessing the Pro51Gly mutation are consistently inferior binders.

Original languageEnglish
Pages (from-to)6951-6960
Number of pages10
JournalBiochemistry
Volume54
Issue number46
DOIs
Publication statusPublished - Oct 28 2015

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Hinges
Mutation
Binding Sites
Oligomerization
Calorimetry
Titration
Dimers
Binders
Monomers
Temperature
cyanovirin N

ASJC Scopus subject areas

  • Biochemistry

Cite this

A Rigid Hinge Region Is Necessary for High-Affinity Binding of Dimannose to Cyanovirin and Associated Constructs. / Li, Zhen; Bolia, Ashini; Maxwell, Jason D.; Bobkov, Andrey A.; Ghirlanda, Giovanna; Ozkan, S. Banu; Margulis, Claudio J.

In: Biochemistry, Vol. 54, No. 46, 28.10.2015, p. 6951-6960.

Research output: Contribution to journalArticle

Li, Zhen ; Bolia, Ashini ; Maxwell, Jason D. ; Bobkov, Andrey A. ; Ghirlanda, Giovanna ; Ozkan, S. Banu ; Margulis, Claudio J. / A Rigid Hinge Region Is Necessary for High-Affinity Binding of Dimannose to Cyanovirin and Associated Constructs. In: Biochemistry. 2015 ; Vol. 54, No. 46. pp. 6951-6960.
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