PMID- 35362254
OWN - NLM
STAT- Publisher
LR  - 20240220
IS  - 1096-9888 (Electronic)
IS  - 1076-5174 (Print)
IS  - 1076-5174 (Linking)
VI  - 57
IP  - 4
DP  - 2022 Mar 14
TI  - A mass spectrometry-based non-radioactive differential radial capillary action of 
      ligand assay (DRaCALA) to assess ligand binding to proteins.
PG  - e4822
LID - 10.1002/jms.4822 [doi]
LID - e4822
AB  - Binding of ligands to macromolecules changes their physicochemical and enzymatic 
      characteristics. Cyclic di-GMP is a second messenger involved in 
      motility/sessility and acute/chronic infection life style transition. Although 
      the GGDEF domain, predominantly a diguanylate cyclase, represents one of the most 
      abundant bacterial domain superfamilies, the number of cyclic di-GMP receptors 
      falls short. To facilitate screening for cyclic di-nucleotide binding proteins, 
      we describe a non-radioactive, matrix-assisted laser desorption and ionization 
      time-of-flight (MALDI-TOF)-based modification of the widely applied differential 
      radial capillary action of ligand assay (DRaCALA). The results of this assay 
      suggest that the diguanylate cyclase/phosphodiesterase variant YciR(Fec101) , but 
      not selected catalytic mutants, bind cyclic di-GMP. HIGHLIGHTS: Cyclic 
      di-nucleotides are ubiquitous second messengers in bacteria. However, few 
      receptors have been identified. Previous screening of cell lysates by 
      differential radial capillary action of ligand assay (DRaCALA) using radioactive 
      ligand identified cyclic di-nucleotide binding proteins. A MALDI-TOF-based 
      DRaCALA was developed to detect cyclic di-nucleotide binding as a non-radioactive 
      alternative. Known cyclic di-GMP binding proteins were verified and potential 
      cyclic di-GMP binding proteins were identified.
CI  - (c) 2022 The Authors. Journal of Mass Spectrometry published by John Wiley & Sons 
      Ltd.
FAU - Cimdins-Ahne, Annika
AU  - Cimdins-Ahne A
AUID- ORCID: 0000-0001-7414-9914
AD  - Department of Microbiology, Tumor and Cell Biology, Biomedicum, Karolinska 
      Institutet, Solna, Sweden.
FAU - Chernobrovkin, Alexey
AU  - Chernobrovkin A
AD  - Department of Medical Biochemistry and Biophysics, Biomedicum, Karolinska 
      Institutet, Solna, Sweden.
AD  - Pelago Bioscience AB, Solna, Sweden.
FAU - Kim, Soo-Kyoung
AU  - Kim SK
AD  - Department of Cell Biology and Molecular Genetics, University of Maryland, 
      College Park, Maryland, USA.
FAU - Lee, Vincent T
AU  - Lee VT
AUID- ORCID: 0000-0002-3593-0318
AD  - Department of Cell Biology and Molecular Genetics, University of Maryland, 
      College Park, Maryland, USA.
FAU - Zubarev, Roman A
AU  - Zubarev RA
AUID- ORCID: 0000-0001-9839-2089
AD  - Department of Medical Biochemistry and Biophysics, Biomedicum, Karolinska 
      Institutet, Solna, Sweden.
AD  - Department of Pharmacological and Technological Chemistry, I.M. Sechenov First 
      Moscow State Medical University, Moscow, Russia.
FAU - Romling, Ute
AU  - Romling U
AUID- ORCID: 0000-0003-3812-6621
AD  - Department of Microbiology, Tumor and Cell Biology, Biomedicum, Karolinska 
      Institutet, Solna, Sweden.
LA  - eng
GR  - Karolinska Institutet/
GR  - 621-2013-4809/Swedish Research Council for Natural Sciences and Engineering/
GR  - CI 239/1-1/Deutsche Forschungsgemeinschaft/
GR  - CI 239/2-1/Deutsche Forschungsgemeinschaft/
PT  - Journal Article
DEP - 20220314
PL  - England
TA  - J Mass Spectrom
JT  - Journal of mass spectrometry : JMS
JID - 9504818
SB  - IM
PMC - PMC9285882
OTO - NOTNLM
OT  - biofilm
OT  - cyclic di-GMP
OT  - diguanylate cyclase/phosphodiesterase
OT  - mass spectrometry
OT  - non-radioactive DRaCALA
EDAT- 2022/04/02 06:00
MHDA- 2022/04/02 06:00
PMCR- 2022/07/15
CRDT- 2022/04/01 05:40
PHST- 2022/02/13 00:00 [received]
PHST- 2022/03/07 00:00 [accepted]
PHST- 2022/04/01 05:40 [entrez]
PHST- 2022/04/02 06:00 [pubmed]
PHST- 2022/04/02 06:00 [medline]
PHST- 2022/07/15 00:00 [pmc-release]
AID - JMS4822 [pii]
AID - 10.1002/jms.4822 [doi]
PST - aheadofprint
SO  - J Mass Spectrom. 2022 Mar 14;57(4):e4822. doi: 10.1002/jms.4822.