The uptake of OxLDLs (oxidized low density lipoproteins) by CD36-expressing macrophages in the arterial intima and the subsequent ‘foam cell’ formation represents a crucial step in the initiation and development of atherosclerotic plaques. The present study has addressed the function of the CD36 N-terminal cytoplasmic domain in the binding and internalization of OxLDL. A selection of CD36 N-terminal cytoplasmic domain mutants were generated and stably expressed in HEK-293 (human embryonic kidney) cells. The capacity of three mutants [CD36_C3/7-A (CD36-C3A/C7A), CD36_D4/R5-A (CD36-D4A/R5A) and CD36_nCPD− (CD36 lacking the N-terminal cytoplasmic domain)] to bind and endocytose OxLDL was then studied using immunofluorescence microscopy and quantitative fluorimetry. Each of the CD36 constructs was expressed at differing levels at the cell surface, as measured by flow cytometry and Western blotting. Following incubation with DiI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate)–OxLDL, cells bearing the CD36_wt (wild-type CD36), CD36_C3/7-A, CD36_D4/R5-A and CD36_nCPD− constructs all internalized DiI–OxLDL into endosomal structures, whereas empty-vector-transfected cells failed to do so, indicating that, unlike the C-terminal cytoplasmic domain, the N-terminal cytoplasmic domain is not essential for the endocytosis of OxLDL. In conclusion, the uptake of OxLDL by CD36 is not reliant on the presence of the CD36 N-terminal cytoplasmic domain. However, the N-terminal cytoplasmic domain may conceivably be implicated in the maturation of CD36.
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July 30 2008
CD36 N-terminal cytoplasmic domain is not required for the internalization of oxidized low-density lipoprotein
Chris McDermott-Roe;
Chris McDermott-Roe
1
*Functional Genomics and Atherothrombosis Unit, Thrombosis Research Institute, London SW3 6LR, U.K.
†Cardiovascular Division, New Hunt's House, Guy's Campus, King's College London, London SE1 1UL, U.K.
1To whom correspondence should be addressed ([email protected]).
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Juliette Martin;
Juliette Martin
*Functional Genomics and Atherothrombosis Unit, Thrombosis Research Institute, London SW3 6LR, U.K.
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Sophie Collot-Teixeira;
Sophie Collot-Teixeira
*Functional Genomics and Atherothrombosis Unit, Thrombosis Research Institute, London SW3 6LR, U.K.
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John L. McGregor
John L. McGregor
*Functional Genomics and Atherothrombosis Unit, Thrombosis Research Institute, London SW3 6LR, U.K.
†Cardiovascular Division, New Hunt's House, Guy's Campus, King's College London, London SE1 1UL, U.K.
‡INSERM Unit 689, Hôpital Lariboisière, Paris, France
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Publisher: Portland Press Ltd
Received:
May 30 2008
Revision Received:
June 16 2008
Accepted:
June 18 2008
Accepted Manuscript online:
June 18 2008
Online ISSN: 1573-4935
Print ISSN: 0144-8463
© The Authors Journal compilation © 2008 Biochemical Society
2008
Biosci Rep (2008) 28 (3): 145–151.
Article history
Received:
May 30 2008
Revision Received:
June 16 2008
Accepted:
June 18 2008
Accepted Manuscript online:
June 18 2008
Connected Content
A commentary has been published:
CD36 N-terminal cytoplasmic domain is not required for the internalization of oxidized low-density lipoproten
Citation
Chris McDermott-Roe, Juliette Martin, Sophie Collot-Teixeira, John L. McGregor; CD36 N-terminal cytoplasmic domain is not required for the internalization of oxidized low-density lipoprotein. Biosci Rep 1 June 2008; 28 (3): 145–151. doi: https://doi.org/10.1042/BSR20080045
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