Molecular organization of human Ficolin-2.
Human Ficolin-2 (L-Ficolin) is an oligomeric serum protein consisting of a collagen-like stalk and fibrinogen-like recognition domains. The protein binds to arrays of sugars present on different microorganisms, enhances phagocytosis and promotes activation of the lectin complement pathway. So far the detailed oligomeric structure and composition of human Ficolin-2 has not been determined. Recombinant human Ficolin-2 was expressed in Chinese hamster ovary cells and its structure and biological functions were investigated by gel filtration, sucrose density gradient ultracentrifugation, mass spectrometry and surface plasmon resonance spectroscopy. It was revealed that Ficolin-2 has a high molecular weight due to extensive disulfide bridge formation. It was able to bind to different ligands, interact with mannose-binding lectin associated serine proteases and activate the complement system. Mass values of 807 and 403 kDa were determined corresponding to a 24-mer and a 12-mer of 34.4 kDa polypeptides. However, the 24-mer was unstable and the 12-mer is likely the major functional form of the protein. Our results are consistent with the view that Ficolin-2 is built up by a mixture of stable homodimers and homotrimers. Based on our findings we propose a model in which disulfide bridges located in the N-terminal region of the polypeptides explain the oligomerization pattern of human Ficolin-2.
Hummelshoj
Tina
T
Tissue Typing Laboratory, Department of Clinical Immunology, Copenhagen University Hospital, Rigshospitalet, 2100 Copenhagen, Denmark.
Thielens
Nicole M
NM
Madsen
Hans O
HO
Arlaud
Gérard J
GJ
Sim
Robert B
RB
Garred
Peter
P
D000818:Animals; D016466:CHO Cells; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003165:Complement System Proteins / Q000737:chemistry; D006224:Cricetinae; D006801:Humans; D037102:Lectins / Q000737:chemistry* / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship; D000097604:Ficolins
D003165:Complement System Proteins; D037102:Lectins; D011994:Recombinant Proteins; D000097604:Ficolins