# Functional insights into the photoactive yellow protein family from homologs, multidomain proteins, and inferred pyp operons

## TL;DR

This paper explores how the PYP protein in bacteria interacts with other proteins to control responses like phototaxis and biofilm formation.

## Contribution

The study identifies candidate proteins involved in PYP signal transduction using bioinformatics and experimental validation.

## Key findings

- PYP homologs and multi-domain proteins suggest functional partners like histidine kinase and methyl accepting chemotaxis proteins.
- Predicted pyp operons include proteins related to c-di-GMP and biofilm formation, such as GGDEF and EAL domains.
- An overexpressed PYP domain from Nitrincola alkalilacustris showed a 447 nm absorbance maximum and a 0.5 s photocycle rate.

## Abstract

Photoactive Yellow Protein (PYP) is a model system for functional protein dynamics and a prototype of the PAS domain superfamily. It is a bacterial photoreceptor that triggers a range of responses in different bacteria: phototaxis, biosynthesis of photo-protective pigments, and light regulation of biofilm formation. An important gap in knowledge on PYP is the signal transduction chain that guides the initial signal from the photoreceptor to various biological responses. Here, we present an expanded set of 984 PYP homologs, providing information on sequence conservation and variation. We analyze this set of PYPs using two bioinformatics approaches to identify candidate proteins that are functionally related to PYP. First, we identified 153 multi-domain proteins containing PYP and analyzed the domain composition of these proteins. Specific preferences for N- or C-terminal placement of the PYP domain were observed. Second, we identified 113 predicted multi-gene operons containing the pyp gene. These two approaches yielded multiple candidates for proteins in the signal transduction chain associated with PYP, particularly histidine kinase (implying phosphorylation), methyl accepting chemotaxis protein (implying phototaxis), and GGDEF and EAL proteins (implying a role of c-di-GMP and biofilm formation). Some of these candidates were present only in multi-domain proteins and others only in pyp operons. Overexpression of the PYP domain from the MCP-fusion protein from Nitrincola alkalilacustris yielded a protein with an absorbance maximum of 447 nm and an overall photocycle rate of 0.5 s. Our results provide a clear basis for future experimental work on identifying signal transduction partners of PYP.