A continuation of structural investigations on expressed domains from Complement Receptor CR2 is proposed. Complement receptor type 2 (CR2, CD21) normally serves as a receptor for complement C3 activation fragments and the immuno-modulatory protein CD23. Expression by B lymphocytes is required for the development of a normal humoral immune response to foreign antigens. This is due to the ability of CR2 to functionally link the binding of C3 fragments iC3b and C3d,g with antigen to the signal transducing capability of the B lymphocyte protein CD19. In addition the Epstein-Bar virus (EBV) utilizes CR2 as its primary means of infecting cells. The investigators along with other groups have previously studied structure-function relationships that govern the interaction of CR2 with the above ligands using strategies involving mutagenesis, and antibody binding. They have found that a domain composed of two 60-70 amino acid segments called N-terminal short consensus repeats (SCRs) contain the only two binding sites for C3 fragments and the EBV protein gp350/220, and contain one of the two sites for CD23. A model for this domain of CR2 and the location of the CD3 binding sites has been created, but the actual three dimensional structure is not known. The investigators propose to apply an NMR method for determining the three dimensional structure of the CR2 domain. They propose to further characterize ligand binding sites using mutagenesis and inhibitor techniques. They also propose to determine how this domain interacts with other regions of CR2. More specifically, their three specific aims are: 1)to determine the structure at high resolution of the human CR2 SCR 1-2 domain. 2)to define the ligand binding sites in the human CR2 SCR 1-2 domain for C3 activation fragments iC3b and C3d,g in addition to binding sites for EBV gp350/220 and CD23. 3) to establish the 3-dimensional solution phase structure of a longer CR2 region containing the SCR 1-2 domain and determine whether the orientation of this domain is influenced by its carboxy-terminal flanking SCRs.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA053615-08
Application #
2907619
Study Section
Special Emphasis Panel (ZRG1-EI (02))
Program Officer
Mccarthy, Susan A
Project Start
1991-07-01
Project End
2002-06-30
Budget Start
1999-07-01
Budget End
2000-06-30
Support Year
8
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Colorado Denver
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045
Kovacs, James M; Hannan, Jonathan P; Eisenmesser, Elan Z et al. (2010) Biophysical investigations of complement receptor 2 (CD21 and CR2)-ligand interactions reveal amino acid contacts unique to each receptor-ligand pair. J Biol Chem 285:27251-8
Quan, Timothy E; Roman, Robert M; Rudenga, Benjamin J et al. (2010) Epstein-Barr virus promotes interferon-alpha production by plasmacytoid dendritic cells. Arthritis Rheum 62:1693-701
Ricklin, Daniel; Tzekou, Apostolia; Garcia, Brandon L et al. (2009) A molecular insight into complement evasion by the staphylococcal complement inhibitor protein family. J Immunol 183:2565-74
Kovacs, James M; Hannan, Jonathan P; Eisenmesser, Elan Z et al. (2009) Mapping of the C3d ligand binding site on complement receptor 2 (CR2/CD21) using nuclear magnetic resonance and chemical shift analysis. J Biol Chem 284:9513-20
Li, Keying; Okemefuna, Azubuike I; Gor, Jayesh et al. (2008) Solution structure of the complex formed between human complement C3d and full-length complement receptor type 2. J Mol Biol 384:137-50
Young, Kendra A; Herbert, Andrew P; Barlow, Paul N et al. (2008) Molecular basis of the interaction between complement receptor type 2 (CR2/CD21) and Epstein-Barr virus glycoprotein gp350. J Virol 82:11217-27
Twohig, Jason; Kulik, Liudmila; Haluszczak, Catherine et al. (2007) Defective B cell ontogeny and immune response in human complement receptor 2 (CR2, CD21) transgenic mice is partially recovered in the absence of C3. Mol Immunol 44:3434-44
Kulik, Liudmila; Marchbank, Kevin J; Lyubchenko, Taras et al. (2007) Intrinsic B cell hypo-responsiveness in mice prematurely expressing human CR2/CD21 during B cell development. Eur J Immunol 37:623-33
Young, Kendra A; Chen, Xiaojiang S; Holers, V Michael et al. (2007) Isolating the Epstein-Barr virus gp350/220 binding site on complement receptor type 2 (CR2/CD21). J Biol Chem 282:36614-25
Ho, Jason; Moir, Susan; Kulik, Liudmila et al. (2007) Role for CD21 in the establishment of an extracellular HIV reservoir in lymphoid tissues. J Immunol 178:6968-74

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