title: Cloning, expression, and purification of the N-terminal heme-binding domain of globin-coupled sensors.
abstract: Globin-coupled sensors (GCSs) are multidomain proteins, consisting of an N-terminal globin domain fused to a variety of C-terminal transmitter domains. Functional classification of GCSs is based on the transmitter domain(s) they possess, broadly falling under either aerotaxis or gene regulation. This chapter describes methods and strategies for cloning, expression, and purification of GCSs for spectroscopic analysis and determination of the minimum heme-binding domain, together with bioinformatic approaches for database searching and examination of domain architectures.
authors.affiliation: Department of Microbiology, University of Hawaii, Honolulu, Hawaii.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Saito
authors.forename: Tracey Allen K
authors.initials: TA
authors.lastname: Freitas
authors.forename: Maqsudul
authors.initials: M
authors.lastname: Alam
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D005914:Globins / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D006420:Hemeproteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary / Q000235:genetics
chemical_list: D006420:Hemeproteins; D006418:Heme; D005914:Globins