Universidad Complutense de Madrid
E-Prints Complutense

The C-terminal Domains of Two Homologous Oleaceae β-1,3-Glucanases Recognize Carbohydrates Differently: Laminarin Binding by NMR

Impacto

Downloads

Downloads per month over past year



Bruix,, Marta and Zamora-Carreras, Héctor and Torres, María and Villalba, Mayte (2015) The C-terminal Domains of Two Homologous Oleaceae β-1,3-Glucanases Recognize Carbohydrates Differently: Laminarin Binding by NMR. Archives of Biochemistry and Biophysics, 580 . pp. 93-101. ISSN 0003-9861

[img]
Preview
PDF
5MB

Official URL: http://www.sciencedirect.com/science/article/pii/S0003986115300084



Abstract

Ole e 9 and Fra e 9 are two allergenic β-1,3-glucanases from olive and ash tree pollens, respectively. Both proteins present a modular structure with a catalytic N-terminal domain and a carbohydrate-binding module (CBM) at the C-terminus. Despite their significant sequence resemblance, they differ in some functional properties, such as their catalytic activity and the carbohydrate-binding ability. Here, we have studied the different capability of the recombinant C-terminal domain of both allergens to bind laminarin by NMR titrations, binding assays and ultracentrifugation. We show that rCtD-Ole e 9 has a higher affinity for laminarin than rCtD-Fra e 9. The complexes have different exchange regimes on the NMR time scale in agreement with the different affinity for laminarin observed in the biochemical experiments. Utilizing NMR chemical shift perturbation data, we show that only one side of the protein surface is affected by the interaction and that the binding site is located in the inter-helical region between α1 and α2, which is buttressed by aromatic side chains. The binding surface is larger in rCtD-Ole e 9 which may account for its higher affinity for laminarin relative to rCtD-Fra e 9.


Item Type:Article
Uncontrolled Keywords:Fra e 9; Ole e 9; Allergy; Ash pollen; β-1,3-Glucanase; Carbohydrate-binding protein; NMR
Subjects:Sciences > Chemistry > Biochemistry
Medical sciences > Medicine > Allergy
ID Code:33120
Deposited On:14 Sep 2015 12:07
Last Modified:01 Sep 2016 23:01

Origin of downloads

Repository Staff Only: item control page