PMID- 33900431
OWN - NLM
STAT- MEDLINE
DCOM- 20211206
LR  - 20211214
IS  - 1432-1017 (Electronic)
IS  - 0175-7571 (Linking)
VI  - 50
IP  - 3-4
DP  - 2021 May
TI  - Dps-DNA interaction in Marinobacter hydrocarbonoclasticus protein: effect of a 
      single-charge alteration.
PG  - 513-521
LID - 10.1007/s00249-021-01538-0 [doi]
AB  - DNA-binding proteins from starved cells (Dps) are members of the ferritin family 
      of proteins found in prokaryotes, with hollow rounded cube-like structures, 
      composed of 12 equal subunits. These protein nanocages are bifunctional enzymes 
      that protect the cell from the harmful reaction of iron and peroxide (Fenton 
      reaction), thus preventing DNA damage by oxidative stress. Ferrous ions are 
      oxidized at specific iron-binding sites in the presence of the oxidant and stored 
      in its cavity that can accommodate up to ca. 500 iron atoms. DNA-binding 
      properties of Dps are associated with the N-terminal, positive charge rich, 
      extensions that can promote DNA binding and condensation, apparently by a 
      cooperative binding mechanism. Here, we describe the binding and protection 
      activities of Marinobacter hydrocarbonoclasticus Dps using Electrophoretic 
      Mobility Shift Essays (EMSA), and synchrotron radiation circular dichroism (SRCD) 
      spectroscopy. While no DNA condensation was observed in the tested conditions, it 
      was possible to determine a Dps-DNA complex formation with an apparent 
      dissociation constant of  6.0 +/- 1.0 microM and a Hill coefficient of 1.2 +/- 0.1. This 
      interaction is suppressed by the inclusion of a single negative charge in the 
      N-terminal region by point mutation. In Dps proteins containing a ferric mineral 
      core (above 96 Fe/protein), DNA binding was impaired. SRCD data clearly showed 
      that no significant modification existed either in secondary structure or protein 
      stability of WT, Q14E variant and core containing proteins. It was, however, 
      interesting to note that, in our experimental conditions, thermal denaturation 
      induced protein aggregation that caused artifacts in thermal denaturation curves, 
      which were dependent on radiation flux and vertical arrangement of the CD cell.
FAU - Jacinto, Joao P
AU  - Jacinto JP
AD  - Molecular Biophysics Laboratory, UCIBIO, Departamento de Quimica, Faculdade de 
      Ciencias e Tecnologia, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal.
FAU - Penas, Daniela
AU  - Penas D
AD  - Molecular Biophysics Laboratory, UCIBIO, Departamento de Quimica, Faculdade de 
      Ciencias e Tecnologia, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal.
FAU - Guerra, Joao P L
AU  - Guerra JPL
AD  - Molecular Biophysics Laboratory, UCIBIO, Departamento de Quimica, Faculdade de 
      Ciencias e Tecnologia, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal.
FAU - Almeida, Ana V
AU  - Almeida AV
AD  - Molecular Biophysics Laboratory, UCIBIO, Departamento de Quimica, Faculdade de 
      Ciencias e Tecnologia, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal.
FAU - Jones, Nykola C
AU  - Jones NC
AD  - ISA, Department of Physics and Astronomy, Aarhus University, 8000, Aarhus C, 
      Denmark.
FAU - Hoffmann, Soren V
AU  - Hoffmann SV
AD  - ISA, Department of Physics and Astronomy, Aarhus University, 8000, Aarhus C, 
      Denmark.
FAU - Tavares, Pedro
AU  - Tavares P
AD  - Molecular Biophysics Laboratory, UCIBIO, Departamento de Quimica, Faculdade de 
      Ciencias e Tecnologia, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal.
FAU - Pereira, Alice S
AU  - Pereira AS
AUID- ORCID: 0000-0001-5567-6073
AD  - Molecular Biophysics Laboratory, UCIBIO, Departamento de Quimica, Faculdade de 
      Ciencias e Tecnologia, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal. 
      masp@fct.unl.pt.
LA  - eng
GR  - PTDC/BIA-PRO/111485/2009/Fundacao para a Ciencia e a Tecnologia/
GR  - PTDC/QUI/64248/2006/Fundacao para a Ciencia e a Tecnologia/
GR  - PD/00193/2012/Fundacao para a Ciencia e a Tecnologia/
GR  - UIDB/04378/2020/Fundacao para a Ciencia e a Tecnologia/
GR  - SFRH/BD/135056/2017/Fundacao para a Ciencia e a Tecnologia/
GR  - PD/BD/135477/2017/Fundacao para a Ciencia e a Tecnologia/
GR  - UIDB/00068/2020/Fundacao para a Ciencia e a Tecnologia/
GR  - PD/BD/135476/2017/Fundacao para a Ciencia e a Tecnologia/
GR  - SFRH/BD/52535/2014/Fundacao para a Ciencia e a Tecnologia/
GR  - CA15126 MOBIEU/COST/
GR  - Agreement 730872/CALIPSOplus/
PT  - Journal Article
DEP - 20210426
PL  - Germany
TA  - Eur Biophys J
JT  - European biophysics journal : EBJ
JID - 8409413
RN  - 0 (Bacterial Proteins)
RN  - 9007-49-2 (DNA)
RN  - E1UOL152H7 (Iron)
RN  - Marinobacter hydrocarbonoclasticus
SB  - IM
MH  - Bacterial Proteins/genetics
MH  - DNA
MH  - Iron
MH  - *Marinobacter
MH  - Models, Molecular
OTO - NOTNLM
OT  - DNA-binding protein from starved cells
OT  - Protein nanocages
OT  - Synchrotron radiation circular dichroism
OT  - Thermal stability
EDAT- 2021/04/27 06:00
MHDA- 2021/12/15 06:00
CRDT- 2021/04/26 12:23
PHST- 2020/10/02 00:00 [received]
PHST- 2021/04/14 00:00 [accepted]
PHST- 2021/04/06 00:00 [revised]
PHST- 2021/04/27 06:00 [pubmed]
PHST- 2021/12/15 06:00 [medline]
PHST- 2021/04/26 12:23 [entrez]
AID - 10.1007/s00249-021-01538-0 [pii]
AID - 10.1007/s00249-021-01538-0 [doi]
PST - ppublish
SO  - Eur Biophys J. 2021 May;50(3-4):513-521. doi: 10.1007/s00249-021-01538-0. Epub 
      2021 Apr 26.