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The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase
- Source :
- Journal of lipid research, vol 61, iss 10, Journal of Lipid Research, Vol 61, Iss 10, Pp 1347-1359 (2020), Luz, J, Beigneux, A P, Asamoto, D K, He, C, Song, W, Allan, C M, Morales, J E, Tu, Y, Kwok, A, Cottle, T, Meiyappan, M, Fong, L G, Kim, J, Ploug, M, Young, S G & Birrane, G 2020, ' The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase ', Journal of Lipid Research, vol. 61, no. 10, pp. 1347-1359 . https://doi.org/10.1194/jlr.RA120000993, J Lipid Res
- Publication Year :
- 2020
- Publisher :
- eScholarship, University of California, 2020.
-
Abstract
- For three decades, the lipoprotein lipase (LPL)-specific monoclonal antibody 5D2 has been used to investigate LPL structure/function and intravascular lipolysis. 5D2 has been used to measure LPL levels, block the triglyceride hydrolase activity of LPL, and prevent the propensity of concentrated LPL preparations to form homodimers. Two early studies on the location of the 5D2 epitope reached conflicting conclusions, but the more convincing report suggested that 5D2 binds to a tryptophan (Trp)-rich loop in the carboxyl terminus of LPL. The same loop had been implicated in lipoprotein binding. Using surface plasmon resonance, we showed that 5D2 binds with high affinity to a synthetic LPL peptide containing the Trp-rich loop of human (but not mouse) LPL. We also showed, by both fluorescence and ultraviolet resonance Raman spectroscopy, that the Trp-rich loop binds lipids. Finally, we used X-ray crystallography to solve the structure of the Trp-rich peptide bound to a 5D2 Fab fragment. The Trp-rich peptide contains a short alpha-helix, with two tryptophans projecting into the antigen recognition site. A proline substitution in the alpha-helix, found in mouse LPL, is expected to interfere with several hydrogen bonds, explaining why 5D2 cannot bind to mouse LPL.
- Subjects :
- 0301 basic medicine
Biochemistry & Molecular Biology
medicine.drug_class
Peptide
QD415-436
030204 cardiovascular system & hematology
Medical Biochemistry and Metabolomics
Monoclonal antibody
Biochemistry
Epitope
Antibodies
03 medical and health sciences
Mice
0302 clinical medicine
Endocrinology
Protein structure
Monoclonal
lipid metabolism
medicine
antibodies
Animals
Humans
Surface plasmon resonance
protein structure
triglycerides
Research Articles
X-ray crystallography
chemistry.chemical_classification
Lipoprotein lipase
Binding Sites
biology
Chemistry
digestive, oral, and skin physiology
Tryptophan
Antibodies, Monoclonal
nutritional and metabolic diseases
Cell Biology
Lipoprotein Lipase
030104 developmental biology
biology.protein
lipids (amino acids, peptides, and proteins)
Biochemistry and Cell Biology
Antibody
Biotechnology
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Journal of lipid research, vol 61, iss 10, Journal of Lipid Research, Vol 61, Iss 10, Pp 1347-1359 (2020), Luz, J, Beigneux, A P, Asamoto, D K, He, C, Song, W, Allan, C M, Morales, J E, Tu, Y, Kwok, A, Cottle, T, Meiyappan, M, Fong, L G, Kim, J, Ploug, M, Young, S G & Birrane, G 2020, ' The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase ', Journal of Lipid Research, vol. 61, no. 10, pp. 1347-1359 . https://doi.org/10.1194/jlr.RA120000993, J Lipid Res
- Accession number :
- edsair.doi.dedup.....d3ee1923dc4ca97066b417156ef0ae5b