scholarly journals Identification of an immunodominant functional domain on human CD36 antigen using human-mouse chimaeric proteins and homologue-replacement mutagenesis

1995 ◽  
Vol 305 (1) ◽  
pp. 221-224 ◽  
Author(s):  
L Daviet ◽  
R Buckland ◽  
M D Puente Navazo ◽  
J L McGregor
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Functional Domain ◽  
Oxidized Low Density Lipoprotein ◽  
Key Residues ◽  
Antibody Epitopes ◽  
Human And Mouse

The human CD36 antigen is a multifunctional membrane glycoprotein that acts as a receptor for thrombospondin, malaria-infected erythrocytes and oxidized low-density lipoprotein, as well as being implicated in the recognition of apoptotic neutrophils by macrophages. OKM5 and other anti-CD36 monoclonal antibodies have been shown to inhibit these CD36 adhesive functions, suggesting that the monoclonal-antibody epitopes and the domains that mediate these events are closely related. Analysis of a series of chimaeric exchanges between human and mouse CD36 shows that six anti-CD36 monoclonal antibodies (OKM5, FA6-152, L103, 5F1, SM phi and 10/5) recognize epitopes within the domain comprising amino acids 155-183. A seventh monoclonal antibody (13/10) binds to another domain that spans amino acids 30-76. Homologue-replacement mutagenesis performed within the human 155-183 immunodominant sequence identifies key residues for the binding of three functional monoclonal antibodies (OKM5, FA6-152 and L103). The fact that antibodies directed against the 155-183 domain can inhibit adhesion suggests that this domain is directly involved in CD36-ligand binding.

A Structural/Functional Domain on Human CD36 Is Involved in the Binding of Anti-Nakα Antibodies

1995 ◽  
Vol 73 (03) ◽  
pp. 543-545 ◽  
Author(s):  
Laurent Daviet ◽  
Marie-Christine Morel-Kopp ◽  
Cécile Kaplan ◽  
John L McGregor
Keyword(s):  
Monoclonal Antibodies ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Functional Domain ◽  
Membrane Glycoprotein ◽  
Low Density ◽  
Tumor Cell Lines ◽  
Oxidized Low Density Lipoprotein ◽  
Functional Region ◽  
Negative Phenotype

SummaryThe human CD36 antigen is an integral membrane glycoprotein expressed by platelets, monocytes, endothelial cells and various tumor cell lines. CD36 acts as a receptor for thrombospondin, collagen, Plasmodium falciparum-infected erythrocytes and oxidized low- density lipoprotein. Individuals possessing the Naka-negative phenotype do not express CD36 and risk developing anti-CD36 isoantibodies upon blood transfusion or during pregnancy. In the present study, we have examined the interaction of an anti-Naka serum with recombinantly expressed CD36. Results obtained show that five functional CD36 monoclonal antibodies (OKM5, FA6-152, L103, ESIV-C7 and 10/5) prevent the binding of the anti-Naka serum whereas a single monoclonal antibody (13/10) has no effect. Consistent with this result, an epitope map of CD36 generated using cross-blocking experiments, indicates that the inhibitory monoclonal antibodies recognize closely- related epitopes whereas 13/10 reacts with a distinct CD36 determinant. Furthermore, we have demonstrated, in a recent study, that OKM5, FA6-152, L103 and 10/5 bind to the same CD36 domain defined by amino acids 155 to 183. Taken together, our results indicate that the 155-183 sequence is important for the binding of the anti-Naka serum to CD36 and may represent a surface-exposed, immunogenic and presumably functional region on human CD36.

scholarly journals The terminal six amino-acids of the carboxy cytoplasmic tail of CD36 contain a functional domain implicated in the binding and capture of oxidized low-density lipoprotein

2002 ◽  
Vol 364 (2) ◽  
pp. 507-515 ◽  
Author(s):  
Eric MALAUD ◽  
Delphine HOURTON ◽  
Louise Marie GIROUX ◽  
Ewa NINIO ◽  
Robin BUCKLAND ◽  
...  
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibody ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Cytoplasmic Tail ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Low Density ◽  
Wild Type ◽  
Oxidized Low Density Lipoprotein

CD36, a major adhesion molecule expressed by monocytes/macrophages, plays a key role in the binding and internalization of oxidized low-density lipoprotein (OxLDL). This adhesion molecule, a member of an important scavenger receptor family, contains a very short C-terminal cytoplasmic tail that is known to induce intracellular signalling events. However, the domains on the cytoplasmic tail involved in such signal transduction are unknown. In this study, we have investigated the functional components of the cytoplasmic tail by site-directed mutagenesis coupled with functional OxLDL and monoclonal antibody (mAb) binding studies. Seven truncated or punctual CD36 constructs, localized in the cytoplasmic tail, were produced by site-directed mutagenesis. Each construct was stably expressed in HEK293 cells. We used a quantitative and a qualitative method, labelling OxLDL with either iodine or rhodamine, to determine the functional importance of the cytoplasmic domains in OxLDL internalization. Results indicate that: (1) a deletion of the last amino-acid (construct K472STOP) significantly reduces, compared with wild-type, the binding, internalization and degradation of OxLDL; (2) truncation of the last six amino-acids (construct R467STOP) significantly reduces OxLDL binding; (3) the above two constructs (K472STOP and R467STOP) showed a reduced rate of OxLDL internalization compared with wild-type; (4) the binding and rate of internalization of an anti-CD36 monoclonal antibody (10/5) was not affected by the above mentioned mutants (K472STOP and R467STOP), compared with wild-type. This study shows, for the first time, a specific site on the CD36 cytoplasmic tail that is critical for the binding, endocytosis and targeting of OxLDL.

Monoclonal antibodies can precipitate low-density lipoprotein. II. Radioimmunoassays with single and combined monoclonal antibodies for determining apolipoprotein B in serum of patients with coronary artery disease.

1985 ◽  
Vol 31 (10) ◽  
pp. 1659-1663 ◽  
Author(s):  
S Marcovina ◽  
B A Kottke ◽  
S J Mao
Keyword(s):  
Coronary Artery Disease ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Coronary Artery ◽  
Apolipoprotein B ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Apo B ◽  
Artery Disease

Abstract We have established four lines of monoclonal antibodies against human low-density lipoproteins (LDL) that, mixed in equal proportions, can precipitate LDL in gel and so can be used for apolipoprotein (apo) B determination in plasma. One monoclonal antibody (clone A), with a relatively low binding affinity to LDL (ka = 0.6 X 10(9) L/mol) and recognizing only two species of apo B, significantly underestimated the concentration of apo B in 74 patients with and 27 without coronary artery disease (CAD). High-affinity monoclonal antibody C (Ka = 3.8 X 10(9) L/mol), which recognized all four apo B species, gave the same value for apo B as determined with the mixture of monoclonal antibodies. The latter results (by radioimmunoassay, y) correlated well with those by radial immunodiffusion (chi): y = 0.994 chi + 0.003 (r = 0.987). The CAD patients showed an increased concentration of apo B as compared to the angiographically documented CAD-negative patients. Except for the values determined by clone B (p = 0.07), the increase was statistically significant (p = 0.002-0.018) for values determined by use of the other clones or their mixture.

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