Interaction of calreticulin with CD40 ligand, TRAIL and Fas ligand.
The molecular chaperone calreticulin has been shown to bind C1q and mannan-binding lectin (MBL), which are constituents of the innate immune defence system. C1q and MBL do not share a large sequence identity but have a similar overall molecular architecture: an N-terminal triple-helical collagen-like domain and a C-terminal globular domain with ligand-binding properties. C1q is a hetero-trimer, while MBL is a homo-trimer, but due to the presence of N-terminal cysteines they both form higher order oligomers of trimers, which are the mature functional molecules. The same molecular architecture is utilized by many other functionally diverse molecules and in this work the interaction of calreticulin with C1q and structurally similar molecules was investigated. In addition to C1q and MBL, CD40 ligand (CD40L), tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas ligand (FasL) were found to bind calreticulin strongly. A low level or no binding was observed for adiponectin, tumour necrosis factor-alpha (TNF-alpha), CD30L, surfactant protein-A and -D and collagen VIII. The interaction with calreticulin required a conformational change in CD40L, TRAIL and FasL and showed the same characteristics as calreticulin's interaction with C1q and MBL: a time-dependent saturable binding to immobilized protein, which was initially sensitive to salt but gradually developed into a salt-insensitive interaction. Thus, the interaction requires a structural change in the interaction partner and leads to a conformational change in calreticulin itself. The implications of these results are that calreticulin may function as a general response modifier for a whole group of immunologically important proteins.
Duus
K
K
Department of Autoimmunology, Statens Serum Institut, Copenhagen, Denmark.
Pagh
R T
RT
Holmskov
U
U
Højrup
P
P
Skov
S
S
Houen
G
G
D052242:Adiponectin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D023201:CD40 Ligand / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D037282:Calreticulin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015922:Complement C1q / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D053222:Fas Ligand Protein / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D020349:Surface Plasmon Resonance; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D052242:Adiponectin; D037282:Calreticulin; D053222:Fas Ligand Protein; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha; D023201:CD40 Ligand; D015922:Complement C1q