-
Je něco špatně v tomto záznamu ?
Negative charge of the AC-to-Hly linking segment modulates calcium-dependent membrane activities of Bordetella adenylate cyclase toxin
A. Sukova, L. Bumba, P. Srb, V. Veverka, O. Stanek, J. Holubova, J. Chmelik, R. Fiser, P. Sebo, J. Masin,
Jazyk angličtina Země Nizozemsko
Typ dokumentu časopisecké články, práce podpořená grantem
- MeSH
- adenylátcyklasový toxin chemie metabolismus MeSH
- AMP cyklický metabolismus MeSH
- biologický transport genetika MeSH
- Bordetella pertussis chemie metabolismus MeSH
- buněčná membrána chemie metabolismus MeSH
- hemolýza genetika MeSH
- konformace proteinů, alfa-helix genetika MeSH
- lidé MeSH
- lipidové dvojvrstvy chemie metabolismus MeSH
- permeabilita buněčné membrány genetika MeSH
- vápník metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Two distinct conformers of the adenylate cyclase toxin (CyaA) appear to accomplish its two parallel activities within target cell membrane. The translocating conformer would deliver the N-terminal adenylyl cyclase (AC) enzyme domain across plasma membrane into cytosol of cells, while the pore precursor conformer would assemble into oligomeric cation-selective pores and permeabilize cellular membrane. Both toxin activities then involve a membrane-interacting 'AC-to-Hly-linking segment' (residues 400 to 500). Here, we report the NMR structure of the corresponding CyaA411-490 polypeptide in dodecylphosphocholine micelles and show that it consists of two α-helices linked by an unrestrained loop. The N-terminal α-helix (Gly418 to His439) remained solvent accessible, while the C-terminal α-helix (His457 to Phe485) was fully enclosed within detergent micelles. CyaA411-490 weakly bound Ca2+ ions (apparent KD 2.6 mM) and permeabilized negatively charged lipid vesicles. At high concentrations (10 μM) the CyaA411-490 polypeptide formed stable conductance units in artificial lipid bilayers with applied voltage, suggesting its possible transmembrane orientation in the membrane-inserted toxin. Mutagenesis revealed that two clusters of negatively charged residues within the 'AC-to-Hly-linking segment' (Glu419 to Glu432 and Asp445 to Glu448) regulate the balance between the AC domain translocating and pore-forming capacities of CyaA in function of calcium concentration.
Citace poskytuje Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc20027866
- 003
- CZ-PrNML
- 005
- 20230519100851.0
- 007
- ta
- 008
- 210105s2020 ne f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1016/j.bbamem.2020.183310 $2 doi
- 035 __
- $a (PubMed)32333856
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a ne
- 100 1_
- $a Sukova, Anna $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic.
- 245 10
- $a Negative charge of the AC-to-Hly linking segment modulates calcium-dependent membrane activities of Bordetella adenylate cyclase toxin / $c A. Sukova, L. Bumba, P. Srb, V. Veverka, O. Stanek, J. Holubova, J. Chmelik, R. Fiser, P. Sebo, J. Masin,
- 520 9_
- $a Two distinct conformers of the adenylate cyclase toxin (CyaA) appear to accomplish its two parallel activities within target cell membrane. The translocating conformer would deliver the N-terminal adenylyl cyclase (AC) enzyme domain across plasma membrane into cytosol of cells, while the pore precursor conformer would assemble into oligomeric cation-selective pores and permeabilize cellular membrane. Both toxin activities then involve a membrane-interacting 'AC-to-Hly-linking segment' (residues 400 to 500). Here, we report the NMR structure of the corresponding CyaA411-490 polypeptide in dodecylphosphocholine micelles and show that it consists of two α-helices linked by an unrestrained loop. The N-terminal α-helix (Gly418 to His439) remained solvent accessible, while the C-terminal α-helix (His457 to Phe485) was fully enclosed within detergent micelles. CyaA411-490 weakly bound Ca2+ ions (apparent KD 2.6 mM) and permeabilized negatively charged lipid vesicles. At high concentrations (10 μM) the CyaA411-490 polypeptide formed stable conductance units in artificial lipid bilayers with applied voltage, suggesting its possible transmembrane orientation in the membrane-inserted toxin. Mutagenesis revealed that two clusters of negatively charged residues within the 'AC-to-Hly-linking segment' (Glu419 to Glu432 and Asp445 to Glu448) regulate the balance between the AC domain translocating and pore-forming capacities of CyaA in function of calcium concentration.
- 650 _2
- $a adenylátcyklasový toxin $x chemie $x metabolismus $7 D037361
- 650 _2
- $a biologický transport $x genetika $7 D001692
- 650 _2
- $a Bordetella pertussis $x chemie $x metabolismus $7 D001886
- 650 _2
- $a vápník $x metabolismus $7 D002118
- 650 _2
- $a buněčná membrána $x chemie $x metabolismus $7 D002462
- 650 _2
- $a permeabilita buněčné membrány $x genetika $7 D002463
- 650 _2
- $a AMP cyklický $x metabolismus $7 D000242
- 650 _2
- $a hemolýza $x genetika $7 D006461
- 650 _2
- $a lidé $7 D006801
- 650 _2
- $a lipidové dvojvrstvy $x chemie $x metabolismus $7 D008051
- 650 _2
- $a konformace proteinů, alfa-helix $x genetika $7 D000072756
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Bumba, Ladislav $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic.
- 700 1_
- $a Srb, Pavel $u Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo namesti. 2, 166 10 Prague, Czech Republic.
- 700 1_
- $a Veverka, Václav, $u Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo namesti. 2, 166 10 Prague, Czech Republic; Department of Cell Biology, Faculty of Science, Charles University, Vinicna 7, 128 00 Prague, Czech Republic. $d 1973- $7 xx0301322
- 700 1_
- $a Stanek, Ondrej $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic.
- 700 1_
- $a Holubova, Jana $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic.
- 700 1_
- $a Chmelik, Josef $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic.
- 700 1_
- $a Fiser, Radovan $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic; Department of Genetics and Microbiology, Faculty of Science, Charles University, Vinicna 5, 128 00 Prague, Czech Republic.
- 700 1_
- $a Sebo, Peter $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic.
- 700 1_
- $a Masin, Jiri $u Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 20 Prague, Czech Republic. Electronic address: masin@biomed.cas.cz.
- 773 0_
- $w MED00000713 $t Biochimica et biophysica acta. Biomembranes $x 1879-2642 $g Roč. 1862, č. 9 (2020), s. 183310
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/32333856 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y a $z 0
- 990 __
- $a 20210105 $b ABA008
- 991 __
- $a 20230519100843 $b ABA008
- 999 __
- $a ok $b bmc $g 1608201 $s 1119046
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2020 $b 1862 $c 9 $d 183310 $e 20200422 $i 1879-2642 $m Biochimica et biophysica acta. Biomembranes $n Biochem Biophys Acta $x MED00000713
- LZP __
- $a Pubmed-20210105
