-
Je něco špatně v tomto záznamu ?
Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics
A. Kadek, D. Kavan, J. Marcoux, J. Stojko, AK. Felice, S. Cianférani, R. Ludwig, P. Halada, P. Man,
Jazyk angličtina Země Nizozemsko
Typ dokumentu časopisecké články, práce podpořená grantem
Odkazy
PubMed
27851982
DOI
10.1016/j.bbagen.2016.11.016
Knihovny.cz E-zdroje
- MeSH
- celobiosa metabolismus MeSH
- cytochromy metabolismus MeSH
- deuterium metabolismus MeSH
- elektrony MeSH
- flaviny metabolismus MeSH
- fungální proteiny metabolismus MeSH
- glykosylace MeSH
- houby metabolismus MeSH
- karbohydrátdehydrogenasy metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- oxygenasy se smíšenou funkcí metabolismus MeSH
- polysacharidy metabolismus MeSH
- proteinové domény MeSH
- proteolýza MeSH
- sekvence aminokyselin MeSH
- statická elektřina MeSH
- transport elektronů fyziologie MeSH
- vodík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. METHODS: To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. RESULTS: HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. CONCLUSIONS: Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. GENERAL SIGNIFICANCE: The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells.
- 000
- 00000naa a2200000 a 4500
- 001
- bmc18016914
- 003
- CZ-PrNML
- 005
- 20190906121806.0
- 007
- ta
- 008
- 180515s2017 ne f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1016/j.bbagen.2016.11.016 $2 doi
- 035 __
- $a (PubMed)27851982
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a ne
- 100 1_
- $a Kadek, Alan $u BioCeV - Institute of Microbiology, The Czech Academy of Sciences, Prumyslova 595, 252 50 Vestec, Czech Republic; Department of Biochemistry, Faculty of Science, Charles University in Prague, Hlavova 8, 128 43 Prague, Czech Republic.
- 245 10
- $a Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics / $c A. Kadek, D. Kavan, J. Marcoux, J. Stojko, AK. Felice, S. Cianférani, R. Ludwig, P. Halada, P. Man,
- 520 9_
- $a BACKGROUND: Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. METHODS: To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. RESULTS: HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. CONCLUSIONS: Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. GENERAL SIGNIFICANCE: The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells.
- 650 _2
- $a sekvence aminokyselin $7 D000595
- 650 _2
- $a karbohydrátdehydrogenasy $x metabolismus $7 D002237
- 650 _2
- $a celobiosa $x metabolismus $7 D002475
- 650 _2
- $a cytochromy $x metabolismus $7 D003580
- 650 _2
- $a deuterium $x metabolismus $7 D003903
- 650 _2
- $a transport elektronů $x fyziologie $7 D004579
- 650 _2
- $a elektrony $7 D004583
- 650 _2
- $a flaviny $x metabolismus $7 D005415
- 650 _2
- $a fungální proteiny $x metabolismus $7 D005656
- 650 _2
- $a houby $x metabolismus $7 D005658
- 650 _2
- $a glykosylace $7 D006031
- 650 _2
- $a vodík $x metabolismus $7 D006859
- 650 _2
- $a koncentrace vodíkových iontů $7 D006863
- 650 _2
- $a oxygenasy se smíšenou funkcí $x metabolismus $7 D006899
- 650 _2
- $a polysacharidy $x metabolismus $7 D011134
- 650 _2
- $a proteinové domény $7 D000072417
- 650 _2
- $a proteolýza $7 D059748
- 650 _2
- $a statická elektřina $7 D055672
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Kavan, Daniel $u BioCeV - Institute of Microbiology, The Czech Academy of Sciences, Prumyslova 595, 252 50 Vestec, Czech Republic; Department of Biochemistry, Faculty of Science, Charles University in Prague, Hlavova 8, 128 43 Prague, Czech Republic.
- 700 1_
- $a Marcoux, Julien $u BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, Université de Strasbourg, 25 rue Becquerel, 67087 Strasbourg, France; IPHC, CNRS, UMR7178, 67087 Strasbourg, France.
- 700 1_
- $a Stojko, Johann $u BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, Université de Strasbourg, 25 rue Becquerel, 67087 Strasbourg, France; IPHC, CNRS, UMR7178, 67087 Strasbourg, France.
- 700 1_
- $a Felice, Alfons K G $u Department of Food Sciences and Technology, BOKU - University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
- 700 1_
- $a Cianférani, Sarah $u BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, Université de Strasbourg, 25 rue Becquerel, 67087 Strasbourg, France; IPHC, CNRS, UMR7178, 67087 Strasbourg, France.
- 700 1_
- $a Ludwig, Roland $u Department of Food Sciences and Technology, BOKU - University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
- 700 1_
- $a Halada, Petr $u BioCeV - Institute of Microbiology, The Czech Academy of Sciences, Prumyslova 595, 252 50 Vestec, Czech Republic.
- 700 1_
- $a Man, Petr $u BioCeV - Institute of Microbiology, The Czech Academy of Sciences, Prumyslova 595, 252 50 Vestec, Czech Republic; Department of Biochemistry, Faculty of Science, Charles University in Prague, Hlavova 8, 128 43 Prague, Czech Republic. Electronic address: pman@biomed.cas.cz.
- 773 0_
- $w MED00000717 $t Biochimica et biophysica acta. G, General subjects $x 0304-4165 $g Roč. 1861, č. 2 (2017), s. 157-167
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/27851982 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y a $z 0
- 990 __
- $a 20180515 $b ABA008
- 991 __
- $a 20190906122139 $b ABA008
- 999 __
- $a ok $b bmc $g 1300538 $s 1013754
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2017 $b 1861 $c 2 $d 157-167 $e 20161113 $i 0304-4165 $m Biochimica et biophysica acta. G, General subjects $n Biochem Biophys Acta $x MED00000717
- LZP __
- $a Pubmed-20180515