title: Antibody fragments for stabilization and crystallization of G protein-coupled receptors and their signaling complexes.
abstract: G protein-coupled receptors (GPCRs) are one of the key players in extracellular signal recognition and their subsequent communications with cellular signaling machinery. Crystallization and high-resolution structure determination of GPCRs has been one of the major advances in the area of GPCR biology over the last 7-8 years. There have primarily been three approaches to GPCR crystallization till date. These are fusion protein strategy, thermostabilization, and antibody fragment-mediated crystallization. Of these, antibody fragment-mediated crystallization has not only provided the first breakthrough in structure determination of a non-rhodopsin GPCR but it has also assisted in obtaining structures of fully active conformations of GPCRs. Antibody fragment approach has also been crucial in obtaining structural information on GPCR signaling complexes. Here, we highlight the specific examples of GPCR crystal structures that have utilized antibody fragments for promoting crystallogenesis and structure solution. We also discuss emerging powerful technologies such as the nanobody technology and the synthetic phage display libraries in the context of GPCR crystallization and underline how these tools are likely to propel key GPCR structural studies in future.
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India. Electronic address: arshukla@iitk.ac.in.
authors.forename: Arun K
authors.initials: AK
authors.lastname: Shukla
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
authors.forename: Charu
authors.initials: C
authors.lastname: Gupta
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Srivastava
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
authors.forename: Deepika
authors.initials: D
authors.lastname: Jaiman
mesh_terms: D000818:Animals; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D008958:Models, Molecular; D019151:Peptide Library; D011487:Protein Conformation; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*; D015398:Signal Transduction; D061905:Single-Domain Antibodies / Q000737:chemistry
chemical_list: D007128:Immunoglobulin Fragments; D019151:Peptide Library; D043562:Receptors, G-Protein-Coupled; D061905:Single-Domain Antibodies
keywords: Antibody fragments; Crystallography; GPCR; Nanobody; Phage display; Signaling