title: Regulation of the Ppr histidine kinase by light-induced interactions between its photoactive yellow protein and bacteriophytochrome domains.
abstract: Ppr is a unique bacteriophytochrome that bleaches rather than forming a far-red-shifted Pfr state upon red light activation. Ppr is also unusual in that it has a blue light photoreceptor domain, PYP, which is N-terminally fused to the bacteriophytochrome domain (Bph). When both photoreceptors are activated by light, the fast phase of Bph recovery (1 min lifetime) corresponds to the formation of an intramolecular long-lived complex between the activated PYP domain and the Bph domain (lifetime of 2-3 days). Since this state is unusually long-lived as compared to other intermediates in the photocycle of both PYP and Bph, we interpret this as formation of a metastable complex between activated PYP and Bph domains that takes days to relax. In the metastable complex, the PYP domain is locked in its activated UV absorbing state and the Bph domain is in a slightly red-shifted state (from 701 to 702 nm), which is photochemically inactive to red or white light. The amount of metastable complex formed increases with the degree of prior activation of PYP, reaching a maximum of 50% when PYP is fully activated compared to 0% when no PYP is activated. The saturation of complex formation at 50% is believed to be due to light-induced heterogeneity within the Ppr dimer. UV irradiation (365 nm) of the metastable complex state photoreverses the activated PYP and the red-shifted Bph to the initial dark state within seconds. We therefore postulate that Ppr functions as a UV-red light sensor and describe the different Ppr states that can be obtained depending on the light quality. Both red and white light upregulate the autokinase activity, while it is downregulated in the dark. The physiological state of Ppr is most likely a mixture of three different states, dark, metastable complex, and red light-activated, with fractional populations whose amounts depend on the light quality of the environment and that regulate the extent of phosphorylation by the kinase.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona, Tucson, Arizona 85721, USA. jkyndt@email.arizona.edu
authors.forename: John A
authors.initials: JA
authors.lastname: Kyndt
authors.forename: John C
authors.initials: JC
authors.lastname: Fitch
authors.forename: Sven
authors.initials: S
authors.lastname: Seibeck
authors.forename: Berthold
authors.initials: B
authors.lastname: Borucki
authors.forename: Maarten P
authors.initials: MP
authors.lastname: Heyn
authors.forename: Terry E
authors.initials: TE
authors.lastname: Meyer
authors.forename: Michael A
authors.initials: MA
authors.lastname: Cusanovich
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D008027:Light; D008954:Models, Biological; D020283:Photoreceptors, Microbial / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000528:radiation effects; D017434:Protein Structure, Tertiary; D013056:Spectrophotometry, Ultraviolet
chemical_list: D001426:Bacterial Proteins; D020283:Photoreceptors, Microbial; C120281:Ppr protein, Rhodospirillum centenum; C050162:photoactive yellow protein, Bacteria