PMID- 28889295
OWN - NLM
STAT- MEDLINE
DCOM- 20180511
LR  - 20180813
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1657
DP  - 2017
TI  - Analysis of c-di-GMP Levels Synthesized by a Photoreceptor Protein in Response to 
      Different Light Qualities Using an In Vitro Enzymatic Assay.
PG  - 187-204
LID - 10.1007/978-1-4939-7240-1_15 [doi]
AB  - Diguanylate cyclases are enzymes that use two GTP molecules to produce one 
      molecule cyclic dimeric guanosine monophosphate (c-di-GMP). This cyclic 
      dinucleotide is an ubiquitous prokaryotic second messenger that controls a 
      variety of cell functions. Several proteins have been described which contain a 
      photoreceptor domain fused to a diguanylate cyclase. The cyanobacterial light 
      sensor Cph2 is responsible for the blue-light induced synthesis of c-di-GMP in 
      Synechocystis sp. PCC 6803. Here, we provide a detailed protocol for an in vitro 
      enzymatic assay with a purified photoreceptor protein using light as the crucial 
      reaction parameter for c-di-GMP synthesis. The assay is accomplished under 
      continuous illumination with light of different quality with inactivation of the 
      enzyme by heat denaturation. Analytics are performed using HPLC-UV.
FAU - Angerer, Veronika
AU  - Angerer V
AD  - Institute of Biology III, Albert-Ludwigs-University Freiburg, Schanzlestr. 1, 
      79104, Freiburg, Germany.
FAU - Essen, Lars-Oliver
AU  - Essen LO
AD  - Department of Chemistry, Philipps-University Marburg, 35032, Marburg, Germany.
FAU - Wilde, Annegret
AU  - Wilde A
AD  - Institute of Biology III, Albert-Ludwigs-University Freiburg, Schanzlestr. 1, 
      79104, Freiburg, Germany. annegret.wilde@biologie.uni-freiburg.de.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Bacterial Proteins)
RN  - 61093-23-0 (bis(3',5')-cyclic diguanylic acid)
RN  - H2D2X058MU (Cyclic GMP)
SB  - IM
MH  - Bacterial Proteins/*metabolism
MH  - Chromatography, High Pressure Liquid
MH  - Cyanobacteria/*physiology/*radiation effects
MH  - Cyclic GMP/*analogs & derivatives/biosynthesis/metabolism
MH  - Enzyme Activation
MH  - Enzyme Assays/methods/standards
MH  - *Light
MH  - Reference Standards
MH  - Spectrophotometry, Ultraviolet
OTO - NOTNLM
OT  - Cyclic diguanylate
OT  - Cyclic nucleotide
OT  - Enzyme-activity assay
OT  - In vitro enzymatic method
OT  - Light-dependent enzymatic activity
OT  - Photosensor
OT  - Photoswitch
OT  - Second messenger
OT  - c-di-GMP
EDAT- 2017/09/11 06:00
MHDA- 2018/05/12 06:00
CRDT- 2017/09/11 06:00
PHST- 2017/09/11 06:00 [entrez]
PHST- 2017/09/11 06:00 [pubmed]
PHST- 2018/05/12 06:00 [medline]
AID - 10.1007/978-1-4939-7240-1_15 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2017;1657:187-204. doi: 10.1007/978-1-4939-7240-1_15.