Polymorphisms in mannan-binding lectin (MBL)-associated serine protease 2 affect stability, binding to MBL, and enzymatic activity

J Immunol. 2009 Mar 1;182(5):2939-47. doi: 10.4049/jimmunol.0802053.

Abstract

Mannan-binding lectin-associated serine protease 2 (MASP-2) is an enzyme of the innate immune system. MASP-2 forms complexes with the pattern recognition molecules mannan-binding lectin (MBL), H-ficolin, L-ficolin, or M-ficolin, and is activated when one of these proteins recognizes microorganisms and subsequently cleaves complement factors C4 and C2, thus initiating the activation of the complement system. Missense polymorphisms of MASP-2 exist in different ethnic populations. To further characterize the nature of these, we have produced and characterized rMASP-2s representing the following naturally occurring polymorphisms: R99Q, D120G, P126L, H155R, 156_159dupCHNH (CHNHdup), V377A, and R439H. Only very low levels of CHNHdup were secreted from the cells, whereas quantities similar to wild-type MASP-2 were found intracellularly, indicating that this mutation results in a misfolded protein. We found that D120G and CHNHdup could not associate with MBL, whereas R99Q, P126L, H155R, V377A, R439H, and wild-type MASP-2 bound equally well to MBL. Accordingly, when D120G and CHNHdup were mixed with MBL, no activation of complement factor C4 was observed, whereas R99Q, P126L, and V377A cleaved C4 with an activity comparable to wild-type MASP-2 and H155R slightly better. In contrast, the R439H variant was deficient in this process despite its normal binding to MBL. This variant was also not able to autoactivate in the presence of MBL and mannan. We find the R439H variant is common in Sub-Saharan Africans with a gene frequency of 10%. Our results indicate that individuals with different types of MASP-2 defects may be identified through genotyping.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution / genetics
  • Amino Acid Substitution / immunology
  • Cell Line
  • Complement C4 / genetics
  • Complement C4 / metabolism
  • Complement Pathway, Mannose-Binding Lectin / genetics*
  • Complement Pathway, Mannose-Binding Lectin / immunology
  • Enzyme Activation / genetics
  • Enzyme Activation / immunology
  • Enzyme Stability / genetics
  • Genetic Predisposition to Disease
  • Genetic Variation / immunology
  • Genotype
  • Humans
  • Mannose-Binding Lectin / metabolism*
  • Mannose-Binding Protein-Associated Serine Proteases / antagonists & inhibitors
  • Mannose-Binding Protein-Associated Serine Proteases / biosynthesis
  • Mannose-Binding Protein-Associated Serine Proteases / genetics*
  • Mannose-Binding Protein-Associated Serine Proteases / metabolism
  • Polymorphism, Single Nucleotide / immunology*
  • Protein Binding / genetics
  • Protein Binding / immunology
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Complement C4
  • Mannose-Binding Lectin
  • Recombinant Proteins
  • MASP2 protein, human
  • Mannose-Binding Protein-Associated Serine Proteases