ArticleThe Biochemical journal2004
Purification and biochemical characterization of the D6 chemokine receptor.
Article in The Biochemical journal, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 71 citations in OpenAlex.
- The Role of Atypical Chemokine Receptor D6 (ACKR2) in Physiological and Pathological Conditions; Friend, Foe, or Both?Frontiers in immunology · 2022Review
- MicroRNA-146 and cell trauma down-regulate expression of the psoriasis-associated atypical chemokine receptor ACKR2.The Journal of biological chemistry · 2018Article
- Mechanisms of Regulation of the Chemokine-Receptor Network.International journal of molecular sciences · 2017Review
- Atypical chemokine receptor ACKR2 controls branching morphogenesis in the developing mammary gland.Development (Cambridge, England) · 2017Article
- Synthesis of Sulfotyrosine-Containing Peptides by Incorporating Fluorosulfated Tyrosine Using an Fmoc-Based Solid-Phase Strategy.Angewandte Chemie (International ed. in English) · 2016Article
- Article
- Chaperoning G protein-coupled receptors: from cell biology to therapeutics.Endocrine reviews · 2014Review
- The N-terminal region of the atypical chemokine receptor ACKR2 is a key determinant of ligand binding.The Journal of biological chemistry · 2014Article
- International Union of Basic and Clinical Pharmacology. [corrected]. LXXXIX. Update on the extended family of chemokine receptors and introducing a new nomenclature for atypical chemokine receptors.Pharmacological reviews · 2014Review
- Overexpression of membrane proteins in mammalian cells for structural studies.Molecular membrane biology · 2013Review
- Milligram production and biological activity characterization of the human chemokine receptor CCR3.PloS one · 2013Article
- One-step immunopurification and lectinochemical characterization of the Duffy atypical chemokine receptor from human erythrocytes.Glycoconjugate journal · 2012Article
- A rapid and efficient way to obtain modified chemokines for functional and biophysical studies.Cytokine · 2011Article
- Atypical chemokine receptors.Experimental cell research · 2011Review
- A recombinant dromedary antibody fragment (VHH or nanobody) directed against human Duffy antigen receptor for chemokines.Cellular and molecular life sciences : CMLS · 2010Article
- Expression, purification and in vitro functional reconstitution of the chemokine receptor CCR1.Protein expression and purification · 2009Article
- BacMam system for high-level expression of recombinant soluble and membrane glycoproteins for structural studies.Protein expression and purification · 2008Article
- Chemokines in and out of the central nervous system: much more than chemotaxis and inflammation.Journal of leukocyte biology · 2008Review
- Location, location, location...site-specific GPCR phosphorylation offers a mechanism for cell-type-specific signalling.Trends in pharmacological sciences · 2008Review
- A novel mechanism for CCR4 in the regulation of macrophage activation in bleomycin-induced pulmonary fibrosis.The American journal of pathology · 2008Article
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
There is much interest in chemokine receptors as therapeutic targets in diseases such as AIDS, autoimmune and inflammatory disorders, and cancer. Hampering such studies is the lack of accurate three-dimensional structural models of these molecules. The CC-chemokine receptor D6 is expressed at exceptionally high levels in heterologous transfectants. Here we report the purification and biochemical characterization of milligram quantities of D6 protein from relatively small cultures of transfected mammalian cells. Importantly, purified D6 retains full functional activity, shown by displaceable binding of 125I-labelled MIP-1beta (macrophage inflammatory protein-1beta) and by complete binding of the receptor to a MIP-1alpha affinity column. In addition, we show that D6 is decorated on the N-terminus by N-linked glycosylation. Mutational analysis reveals that this glycosylation is dispensable for ligand binding and high expression in transfected cells. Metabolic labelling has revealed the receptor to also be sulphated and phosphorylated. Phosphorylation is ligand independent and is not enhanced by ligand binding and internalization, suggesting similarities with the viral chemokine receptor homologue US28. Like US28, an analysis of the full cellular complement of D6 in transfected cells indicates that >80% is found associated with intracellular vesicular structures. This may account for the high quantities of D6 that can be synthesized in these cells. These unusual properties of D6, and the biochemical characterization described here, leads the way towards work aimed at generating the three-dimensional structure of this seven-transmembrane-spanning receptor.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.