Mutational Analysis of the Complement Receptor Type 2 (CR2/CD21)–C3d Interaction Reveals a Putative Charged SCR1 Binding Site for C3d

Hannan, Jonathan P.; Young, Kendra A.; Guthridge, Joel M.; Asokan, Rengasamy; Szakonyi, Gerda; Chen, Xiaojiang S.; Holers, V. Michael

doi:10.1016/j.jmb.2004.12.007

Cited by 45 publications

(87 citation statements)

References 38 publications

(46 reference statements)

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“…However, contrary to suggestions in earlier studies, the x-ray co-crystal structure showed only SCR2 to be in contact with C3d (33). Subsequent amino acid mutagenesis data confirmed the SCR2 x-ray cocrystal contact site as well as providing support for the biochemical data in that multiple amino acids in SCR1 were required for a high affinity C3d interaction (36). The solution structure models of CR2-C3d complexes determined from analytical centrifugation and x-ray/neutron scattering studies also point to both SCR1 and SCR2 interacting with C3d (35).…”

Section: Discussioncontrasting

confidence: 55%

“…Human C3d for peptide discovery, ELISA binding studies, and NMR titrations was generated using the pGEX expression system in E. coli as previously described (36). Briefly, ampicillin-resistant colonies were used to start overnight cultures that were expanded to 1 liter and grown at 37°C until an A 600 of 0.3 was achieved.…”

Section: Methodsmentioning

confidence: 99%

“…The CR2-C3d interaction has been studied, and the results of site-directed mutagenesis suggest that both SCR domains are required for binding and that the interaction is in part charge-mediated (36). Residues found to be required for interaction were Arg 13 , Tyr 16 , Ser 32 , Arg 28 , Arg 36 , Lys 41 , Lys 50 , Lys 57 , Lys 67 , and Arg 83 . Despite substantial progress in understanding CR2-ligand interactions, several key questions remain.…”

mentioning

confidence: 99%

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Mapping of the C3d Ligand Binding Site on Complement Receptor 2 (CR2/CD21) Using Nuclear Magnetic Resonance and Chemical Shift Analysis

Kovacs

Hannan

Eisenmesser

et al. 2009

Journal of Biological Chemistry

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Complement receptor 2 (CR2, CD21) is a cell membrane protein, with 15 or 16 extracellular short consensus repeats (SCRs), that promotes B lymphocyte responses and bridges innate and acquired immunity. The most distally located SCRs (SCR1-2) mediate the interaction of CR2 with its four known ligands (C3d, Epstein-Barr virus gp350, interferon-␣, and CD23). Inhibitory monoclonal antibodies against SCR1-2 block binding of all ligands. To develop ligand-specific inhibitors that would also assist in identifying residues unique to each receptor-ligand interaction, phage were selected from randomly generated libraries by panning with recombinant SCR1-2, followed by specific ligand-driven elution. Derived peptides were tested by competition ELISA. One peptide, C3dp1 (APQHLSSQYSRT) exhibited ligand-specific inhibition at midmicromolar IC 50 . C3d was titrated into 15 N-labeled SCR1-2, which revealed chemical shift changes indicative of specific intermolecular interactions. With backbone assignments made, the chemical shift changes were mapped onto the crystal structure of SCR1-2. With regard to C3d, the binding surface includes regions of SCR1, SCR2, and the inter-SCR linker, specifically residues

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Section: Discussioncontrasting

confidence: 55%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Mapping of the C3d Ligand Binding Site on Complement Receptor 2 (CR2/CD21) Using Nuclear Magnetic Resonance and Chemical Shift Analysis

Kovacs

Hannan

Eisenmesser

et al. 2009

Journal of Biological Chemistry

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“…Moreover, this structure of the complex showed interactions only between CCP2 of CR2 and C3d and no direct interactions between CCP1 of CR2 and C3d. Two recent studies by the same authors showed that CCP1 of CR2 probably contacts C3d directly (Gilbert et al, 2005;Hannan et al, 2005). Although significant structural differences are apparent between TED of C3 and C3d, the CR2 CCP2-binding site is very similar in the two structures.…”

Section: C3b Fragments Signalingmentioning

confidence: 85%

Structural insights into the central complement component C3

Janssen

Gros

2007

Molecular Immunology

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C3 is a central protein of the complement system, which is important to immune defense and provides a link between innate and adaptive immunity. Three pathways of complement activation converge at the activation of C3 yielding a diverse set of biological responses. This versatile and flexible molecule interacts with various proteins to fulfill its functions. Here we review recent insights gained from the crystal structure determinations of human, native C3 and its physiological down-regulation product C3c. The data provided, for the first time, a complete and detailed view of the composition and arrangement of the domains in C3. Comparison of C3 with C3c indicates marked flexibility of the molecule, particularly in the ␣-chain. We discuss the observed domain rearrangements, conformational changes and the location of various protein binding sites. These detailed, and structural, insights are important for developing models of the molecular mechanisms underlying the diverse biological activities of this large and complex molecule.

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“…However, the observed structural arrangement of the complex is controversial. Very recently it was shown that, in addition, the CCP1 domain of CR2 probably makes direct contacts to C3d 125,126 . Nevertheless, the exact site of interaction of CCP1 on C3d still remains unknown.…”

Section: Signaling Roles Of C3b Fragmentsmentioning

confidence: 99%

Conformational Complexity of Complement Component C3

Janssen

Gros

Advances in Experimental Medicine and Biology

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Mutational Analysis of the Complement Receptor Type 2 (CR2/CD21)–C3d Interaction Reveals a Putative Charged SCR1 Binding Site for C3d

Cited by 45 publications

References 38 publications

Mapping of the C3d Ligand Binding Site on Complement Receptor 2 (CR2/CD21) Using Nuclear Magnetic Resonance and Chemical Shift Analysis

Mapping of the C3d Ligand Binding Site on Complement Receptor 2 (CR2/CD21) Using Nuclear Magnetic Resonance and Chemical Shift Analysis

Structural insights into the central complement component C3

Conformational Complexity of Complement Component C3

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