CCR5 internalisation and signalling have different dependence on membrane lipid raft integrity

Cardaba, Clara Moyano; Kerr, Jason S.; Müeller, Anja

doi:10.1016/j.cellsig.2008.05.014

Cited by 33 publications

(36 citation statements)

References 45 publications

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“…We previously showed that depleting the plasma membrane of cholesterol by using methyl--cyclodextrin (MCD) inhibits CCR5 induced calcium release in stably transfected CHO and HEK cell lines, whereas complexing cholesterol with filipin or nystatin does not affect signalling by the receptor [11]. Here we investigate in more Figure 1C, D).This result is a contradiction of our previous data, where MCD treatment significantly blocks the release of calcium [11].…”

Section: Depletion Of Cholesterol Increases Signalling In Thp-1 Ccontrasting

confidence: 93%

“…Here we investigate in more Figure 1C, D).This result is a contradiction of our previous data, where MCD treatment significantly blocks the release of calcium [11].…”

Section: Depletion Of Cholesterol Increases Signalling In Thp-1 Ccontrasting

confidence: 89%

“…Previous reports by us and others highlighted the importance of cholesterol in the plasma membrane for effective CCR5 signalling [6,8,11,22]. Altogether there is a certain problematic with 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 13 specific, since it can be observed in CHO.CCR5 cells, but not in THP-1 cells [24].…”

Section: Discussionmentioning

confidence: 97%

“…We already showed previously that different agents that affect cholesterol in cells have different effect on signalling by receptors [11]. We therefore were interested in understanding how these different compounds affect the overall levels of cholesterol in the cells.…”

Section: As Well As Statins Lower the Amount Of Cholesterol In Thmentioning

confidence: 99%

“…We showed that MCD treatment lead to failure of CCR5 to inhibit forskolin-stimulated accumulation of cAMP and it also changed the potency of CCL3 for intracellular calcium release, however cholesterol depletion had no effect on receptor internalisation. Furthermore, depletion of cholesterol changed the coupling of CCR5 from Gi G-protein to a PTX independent G-protein, as shown by the release of intracellular calcium after PTX treatment [11].…”

Section: Introductionmentioning

confidence: 99%

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Distinct modes of molecular regulation of CCL3 induced calcium flux in monocytic cells

Cardaba

Müeller

2009

Biochemical Pharmacology

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Page 1 of 32A c c e p t e d M a n u s c r i p t 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 by MCD addition, which shows that the cholesterol content in the membrane is only one factor in determining the amount of receptor response. We show that CCR5signalling is dependant on thapsigargin-sensitive Ca 2+ stores and on activation of ryanodine receptors as well as InsP3 receptors or store-operated channels.Cholesterol depletion with MCD changes the thapsigargin sensitivity in THP-1 cells * ManuscriptPage 2 of 32 A c c e p t e d M a n u s c r i p t 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 2 and also alters receptor-G-protein coupling towards pertussis toxin (PTX) independent G-proteins. Cholesterol removal by MCD in THP-1 cells has far reaching consequences for receptor activation and signalling and emphasises the need for a clearer understanding of how membrane fluidity affects receptor signalling events.

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Section: Depletion Of Cholesterol Increases Signalling In Thp-1 Ccontrasting

confidence: 93%

“…Here we investigate in more Figure 1C, D).This result is a contradiction of our previous data, where MCD treatment significantly blocks the release of calcium [11].…”

Section: Depletion Of Cholesterol Increases Signalling In Thp-1 Ccontrasting

confidence: 89%

Section: Discussionmentioning

confidence: 97%

Section: As Well As Statins Lower the Amount Of Cholesterol In Thmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Distinct modes of molecular regulation of CCL3 induced calcium flux in monocytic cells

Cardaba

Müeller

2009

Biochemical Pharmacology

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Stable and sensitive probes for lysosomes: Cell‐penetrating fluorescent calix[4]arenes accumulate in acidic vesicles

et al. 2010

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The uptake of a fluorescently labeled cationic calix [4] (NBDCalAm) in live, nonfixed cells has been investigated. The compound is taken into the cells rapidly and shows distinct endosomal distribution after 2 hours. This distribution pattern shows colocalization with lysosomal staining. The uptake is not altered by inhibition of clathrin or caveolae dependent pathways nor by depletion of the cellular ATP-pool. Immediately after uptake the probe is localized in the Golgi and brefeldin A treatment prevents transport to lysosomes. Pulse chase experiments with bafilomycin A1, monensin, and sodium azide showed that accumulation and retention of the probe in lysosomes is primarily driven by the activity of vacuolar ATPases. The NBD labeled calix[4]arene provides a very stable and sensitive marker for lysosomes, and has a considerable advantage over some commercially available lysosomal markers in so far that the fluorescent signal is stable even when the cells are incubated in dye-free medium after staining. ' 2010 International Society for Advancement of Cytometry Key termslysosome; calixarene; cell-penetration; immunofluorescence CALIXARENES (1) are a family of readily synthesized and functionalized macrocycles which have found applications in a number of research areas. In particular the ability of derivatives to complex anions (2,3) and cations (4,5) has been exploited for the development of sensors. Recently, interest has focused on the use of calixarene derivatives in biological systems be it as drugs, drug delivery systems, or imaging agents. In the design of drug molecules, particular attention has been paid to calixarenes as antimicrobials (6-8), as vaccines (9) and as anticancer agents (10). In the field of drug delivery, agents for nucleic acids based on single calixarenes (11-13) have shown promise and we have recently shown that larger arrays of cationic calix[4]arene, so-called multicalixarenes, are able to transfect DNA effectively (14). In addition a number of studies have combined the ability of calixarene derivatives to complex cations with their low toxicity in the development of noncovalently labeled protein MRI imaging agents (15,16).While the nontoxic (17) and non-immunogenicity (18,19) status of calixarenes has been established, less information is available on how calixarenes are processed within cells and their cellular fate, both of which are important features which must be characterized before such derivatives are taken further for clinical development. We recently demonstrated that fluorescently labeled calix [4]arene have the potential to track progress of the macrocycle in cells, as an NBD labeled cationic derivative was readily and speedily taken up into cells and provided a very stable, nontoxic read-out system to investigate cellular localization (20). The absorbance and emission spectra for the NBDCalAm showed that it can easily be visualized using a GFP filter (20).

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Putative cholesterol‐binding sites in human immunodeficiency virus (HIV) coreceptors CXCR4 and CCR5

2012

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Using molecular docking, we identified a cholesterol-binding site in the groove between transmembrane helices 1 and 7 near the inner membrane-water interface of the G protein-coupled receptor CXCR4, a coreceptor for HIV entry into cells. In this docking pose, the amino group of lysine K67 establishes a hydrogen bond with the hydroxyl group of cholesterol, whereas tyrosine Y302 stacks with cholesterol by its aromatic side chain, and a number of residues form hydrophobic contacts with cholesterol. Sequence alignment showed that a similar putative cholesterol-binding site is also present in CCR5, another HIV coreceptor. We suggest that the interaction of cholesterol with these putative cholesterol-binding sites in CXCR4 and CCR5 is responsible for the presence of these receptors in lipid rafts, for the effect of cholesterol on their conformational stability and function, and for the role that cell cholesterol plays in the cell entry of HIV strains that use these membrane proteins as coreceptors. We propose that mutations of residues that are involved in cholesterol binding will make CXCR4 and CCR5 insensitive to membrane cholesterol content. Cholesterol-binding sites in HIV coreceptors are potential targets for steroid drugs that bind to CXCR4 and CCR5 with higher binding affinity than cholesterol, but do not stabilize the native conformation of these proteins.

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CCR5 internalisation and signalling have different dependence on membrane lipid raft integrity

Cited by 33 publications

References 45 publications

Distinct modes of molecular regulation of CCL3 induced calcium flux in monocytic cells

Distinct modes of molecular regulation of CCL3 induced calcium flux in monocytic cells

Stable and sensitive probes for lysosomes: Cell‐penetrating fluorescent calix[4]arenes accumulate in acidic vesicles

Putative cholesterol‐binding sites in human immunodeficiency virus (HIV) coreceptors CXCR4 and CCR5

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