Cloning, expression, and purification of the N-terminal heme-binding domain of globin-coupled sensors.
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.
Saito
Jennifer A
JA
Department of Microbiology, University of Hawaii, Honolulu, Hawaii.
Freitas
Tracey Allen K
TA
Alam
Maqsudul
M
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
D006420:Hemeproteins; D006418:Heme; D005914:Globins