Flexibility in Mannan-Binding Lectin-Associated Serine Proteases-1 and -2 Provides Insight on Lectin Pathway Activation

Structure. 2017 Feb 7;25(2):364-375. doi: 10.1016/j.str.2016.12.014. Epub 2017 Jan 19.

Abstract

The lectin pathway of complement is activated by complexes comprising a recognition component (mannose-binding lectin, serum ficolins, collectin-LK or collectin-K1) and a serine protease (MASP-1 or MASP-2). MASP-1 activates MASP-2, and MASP-2 cleaves C4 and C4b-bound C2. To clarify activation, new crystal structures of Ca2+-bound MASP dimers were determined, together with their solution structures from X-ray scattering, analytical ultracentrifugation, and atomistic modeling. Solution structures of the CUB1-EGF-CUB2 dimer of each MASP indicate that the two CUB2 domains were tilted by as much as 90° compared with the crystal structures, indicating considerable flexibility at the EGF-CUB2 junction. Solution structures of the full-length MASP dimers in their zymogen and activated forms revealed similar structures that were much more bent than anticipated from crystal structures. We conclude that MASP-1 and MASP-2 are flexible at multiple sites and that this flexibility may permit both intra- and inter-complex activation.

Keywords: X-ray scattering; analytical ultracentrifugation; atomistic modeling; complement; lectin pathway; rat MASP.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • CHO Cells
  • Calcium / chemistry*
  • Calcium / immunology
  • Cations, Divalent
  • Cloning, Molecular
  • Complement Pathway, Mannose-Binding Lectin / genetics*
  • Complement Pathway, Mannose-Binding Lectin / immunology
  • Cricetulus
  • Crystallography, X-Ray
  • Gene Expression
  • Humans
  • Mannose-Binding Protein-Associated Serine Proteases / chemistry*
  • Mannose-Binding Protein-Associated Serine Proteases / genetics
  • Mannose-Binding Protein-Associated Serine Proteases / immunology
  • Models, Molecular
  • Protein Binding
  • Protein Conformation, alpha-Helical
  • Protein Conformation, beta-Strand
  • Protein Interaction Domains and Motifs
  • Rats
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • Cations, Divalent
  • Recombinant Proteins
  • Mannose-Binding Protein-Associated Serine Proteases
  • Masp1 protein, rat
  • Masp2 protein, rat
  • Calcium