Physical methods for studying flavoprotein photoreceptors.

Abstract:

:Molecular mechanisms of dark-to-light state transitions in flavoprotein photoreceptors have been the subject of intense investigation. Blue-light sensing flavoproteins fall into three general classes that share aspects of their activation processes: LOV domains, BLUF proteins, and cryptochromes. In all cases, light-induced changes in flavin redox, protonation, and bonding states result in hydrogen-bond and conformational rearrangements important for regulation of downstream targets. Physical characterization of these flavoprotein states can provide valuable insights into biological function, but clear conclusions are often challenging to draw owing to complexities of data collection and interpretation. In this chapter, we briefly review the three classes of flavoprotein photoreceptors and provide methods for their recombinant production, reconstitution with flavin cofactor, and characterization. We then relate best practices and special considerations for the application of several types of spectroscopies, redox potential measurements, and X-ray scattering experiments to photosensitive flavoproteins. The methods presented are generally accessible to most laboratories.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Yee EF,Chandrasekaran S,Lin C,Crane BR

doi

10.1016/bs.mie.2019.03.023

subject

Has Abstract

pub_date

2019-01-01 00:00:00

pages

509-544

eissn

0076-6879

issn

1557-7988

pii

S0076-6879(19)30104-1

journal_volume

620

pub_type

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