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Crystallization and X‐ray analysis of monodisperse human properdin
Author(s) -
Pedersen Dennis Vestergaard,
Revel Margot,
Gadeberg Trine Amalie Fogh,
Andersen Gregers Rom
Publication year - 2019
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x18018150
Subject(s) - monomer , properdin , cleavage (geology) , chemistry , protease , crystallization , recombinant dna , protein structure , complement system , biophysics , stereochemistry , biochemistry , crystallography , biology , antibody , genetics , enzyme , gene , polymer , organic chemistry , paleontology , fracture (geology)
The 54 kDa protein properdin, also known as factor P (FP), plays a major role in the complement system through the stabilization of the alternative pathway convertases. FP circulates in the blood as cyclic dimers, trimers and tetramers, and this heterogeneity challenges detailed structural insight into the mechanism of convertase stabilization by FP. Here, the generation of an intact FP monomer and a variant monomer with the third thrombospondin repeat liberated is described. Both FP monomers were excised from recombinant full‐length FP containing internal cleavage sites for TEV protease. These FP monomers could be crystallized, and complete data sets extending to 2.8 Å resolution for the intact FP monomer and to 3.5 Å resolution for the truncated variant were collected. The principle of specific monomer excision and domain removal by the insertion of a protease cleavage site may be broadly applicable to structural studies of oligomeric, flexible and modular proteins.