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The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase

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Harvard

Luz, J, Beigneux, AP, Asamoto, DK, He, C, Song, W, Allan, CM, Morales, JE, Tu, Y, Kwok, A, Cottle, T, Meiyappan, M, Fong, LG, Kim, J, Ploug, M, Young, SG & Birrane, G 2020, 'The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase', Journal of Lipid Research, bind 61, nr. 10, s. 1347-1359. https://doi.org/10.1194/jlr.RA120000993

APA

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, 61(10), 1347-1359. https://doi.org/10.1194/jlr.RA120000993

CBE

Luz J, Beigneux AP, Asamoto DK, He C, Song W, Allan CM, Morales JE, Tu Y, Kwok A, Cottle T, Meiyappan M, Fong LG, Kim J, Ploug M, Young SG, Birrane G. 2020. The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase. Journal of Lipid Research. 61(10):1347-1359. https://doi.org/10.1194/jlr.RA120000993

MLA

Vancouver

Author

Luz, John ; Beigneux, Anne P ; Asamoto, DeeAnn K ; He, Cuiwen ; Song, Wenxin ; Allan, Christopher M ; Morales, Jazmin E ; Tu, Yiping ; Kwok, Adam ; Cottle, Thomas ; Meiyappan, Muthuraman ; Fong, Loren G ; Kim, Judy ; Ploug, Michael ; Young, Stephen G ; Birrane, Gabriel. / The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase. I: Journal of Lipid Research. 2020 ; Bind 61, Nr. 10. s. 1347-1359.

Bibtex

@article{ca0e1e86219a4bfc861a5d73320840bf,
title = "The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase",
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.",
keywords = "X-ray crystallography, antibodies, lipid metabolism, protein structure, triglycerides",
author = "John Luz and Beigneux, {Anne P} and Asamoto, {DeeAnn K} and Cuiwen He and Wenxin Song and Allan, {Christopher M} and Morales, {Jazmin E} and Yiping Tu and Adam Kwok and Thomas Cottle and Muthuraman Meiyappan and Fong, {Loren G} and Judy Kim and Michael Ploug and Young, {Stephen G} and Gabriel Birrane",
note = "Published under license by The American Society for Biochemistry and Molecular Biology, Inc.",
year = "2020",
month = jul,
day = "20",
doi = "10.1194/jlr.RA120000993",
language = "English",
volume = "61",
pages = "1347--1359",
journal = "Journal of Lipid Research",
issn = "0022-2275",
publisher = "American Society for Biochemistry and Molecular Biology, Inc",
number = "10",

}

RIS

TY - JOUR

T1 - The structural basis for monoclonal antibody 5D2 binding to the tryptophan-rich loop of lipoprotein lipase

AU - Luz, John

AU - Beigneux, Anne P

AU - Asamoto, DeeAnn K

AU - He, Cuiwen

AU - Song, Wenxin

AU - Allan, Christopher M

AU - Morales, Jazmin E

AU - Tu, Yiping

AU - Kwok, Adam

AU - Cottle, Thomas

AU - Meiyappan, Muthuraman

AU - Fong, Loren G

AU - Kim, Judy

AU - Ploug, Michael

AU - Young, Stephen G

AU - Birrane, Gabriel

N1 - Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

PY - 2020/7/20

Y1 - 2020/7/20

N2 - 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.

AB - 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.

KW - X-ray crystallography

KW - antibodies

KW - lipid metabolism

KW - protein structure

KW - triglycerides

U2 - 10.1194/jlr.RA120000993

DO - 10.1194/jlr.RA120000993

M3 - Journal article

C2 - 32690595

VL - 61

SP - 1347

EP - 1359

JO - Journal of Lipid Research

JF - Journal of Lipid Research

SN - 0022-2275

IS - 10

ER -

ID: 60626901