VOLUME 570. Chemokines

Available here.

"Because of their unusually broad clinical significance, they have been aggressively pursued as therapeutic targets, and the scientific community has realized the importance of understanding the complexities of how these proteins function in order to bring more chemokine-targeted drugs to the clinic. This understanding has been accompanied by increasingly sophisticated methods for studying the molecular and cellular mechanisms of chemokines in normal physiology and disease. It is instructive to look back at the protocols documented in the first Methods of Enzymology volumes from 1997 compared to this volume. While many of these early methods remain as standard laboratory practice, others have been replaced by more efficient methods, or methods that were never feasible before. As documented herein, there are improved ways to carry out assays of binding and signaling that in many cases utilize biosensor or imaging technologies. For more sophisticated assays of cell migration than classic Boyden chambers, microfluidic devices have been developed that allow real-time monitoring of cell migration with control of gradient steepness and flow. Moreover, imaging methods for monitoring migration of specific cell types in in vivo models of disease have been developed. It has become possible not only to make chemokines routinely but also to rapidly generate novel modified chemokines with defined pharmacological properties that hold promise as biologics. Whereas the concept of determining structures of chemokine receptors in complex with their natural ligands and with small molecule antagonists was a dream in 1997, it is now possible, and in fact several such structures have now been solved because of the ability to express and purify receptors. This volume covers these and a wide variety of other methods that I believe will be useful to the chemokine community at large."