Structure–function dissection of D6, an atypical scavenger receptor

Nibbs, R. J.B. , McLean, P., McCulloch, C., Riboldi‐Tunnicliffe, A., Blair, E., Zhu, Y., Isaacs, N. and Graham, G. J. (2009) Structure–function dissection of D6, an atypical scavenger receptor. In: Abelson, J. N. and Simon, M. I. (eds.) Chemokines, Part A. Series: Methods in enzymology (460). Academic Press: Amsterdam, pp. 245-261. ISBN 9780123749086 (doi: 10.1016/S0076-6879(09)05212-4)

Abstract

Chemokines direct leukocyte migration by activating intracellular signalling pathways through G-protein coupled chemokine receptors. However, they also bind to other surface proteins, including a group of molecules which we refer to as ‘atypical’ chemokine receptors. One such molecule is D6. D6 is structurally-related to other chemokine receptors, and binds specific pro-inflammatory chemokines with high affinity, but surprisingly, when expressed in heterologous cell lines, it is unable to transduce signals after chemokine engagement. Instead, by using the approaches outlined in this chapter, evidence has emerged that D6 acts as a chemokine scavenger which uses unique intracellular trafficking properties to continuously sequester extracellular chemokines into cells. It is envisaged that this suppresses inflammation in vivo by limiting pro-inflammatory chemokine bioavailability, and indeed, D6 deficient mice show exaggerated inflammatory responses to a variety of challenges. In addition to the in vitro functional studies, we also describe the methods we have used to express, purify and analyse large quantities of D6 protein. The unusually high stability of D6 and its broad subcellular distribution enables D6 to be expressed to very high levels in transfected cells, making it possible, at least in principal, to produce enough D6 to allow for purification of quantities suitable for crystallisation. This is a key step on the path towards generating a three-dimensional structure of the molecule. Thus, the protocols we outline have helped establish chemokine scavenging as a novel paradigm in chemokine biology, and may also ultimately provide unprecedented insight into the structure of D6 and other chemokine receptors.

• Item Type: Book Sections
• Status: Published
• Glasgow Author(s) Enlighten ID: Blair, Miss Emma and Nibbs, Professor Rob and Isaacs, Professor Neil and McCulloch, Mrs Clare and Zhu, Dr Yanshi and Graham, Professor Gerard
• Authors: Nibbs, R. J.B., McLean, P., McCulloch, C., Riboldi‐Tunnicliffe, A., Blair, E., Zhu, Y., Isaacs, N., and Graham, G. J.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity; College of Science and Engineering > School of Chemistry
• Publisher: Academic Press
• ISSN: 00766879
• ISBN: 9780123749086

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