Integrative structure determination reveals functional global flexibility for an ultra-multimodular arabinanase

Abstract

AbnA is an extracellular GH43 α-L-arabinanase from Geobacillus stearothermophilus, a key bacterial enzyme in the degradation and utilization of arabinan. We present herein its full-length crystal structure, revealing the only ultra-multimodular architecture and the largest structure to be reported so far within the GH43 family. Additionally, the structure of AbnA appears to contain two domains belonging to new uncharacterized carbohydrate-binding module (CBM) families. Three crystallographic conformational states are determined for AbnA, and this conformational flexibility is thoroughly investigated further using the “integrative structure determination” approach, integrating molecular dynamics, metadynamics, normal mode analysis, small angle X-ray scattering, dynamic light scattering, cross-linking, and kinetic experiments to reveal large functional conformational changes for AbnA, involving up to ~100 Å movement in the relative positions of its domains. The integrative structure determination approach demonstrated here may apply also to the conformational study of other ultra-multimodular proteins of diverse functions and structures.

Lansky et al. describe the structural analysis of an ultra-multimodular GH43 arabinanase, AbnA. They discover a new carbohydrate binding module and analyze the large conformational transitions of the enzyme with the integrative structure determination approach, which may be applied to the conformational studies of other ultramultimodular proteins as well.