HydG, the "dangler" iron, and catalytic production of free CO and CN
Journal
Dalton transactions (Cambridge, England : 2003)
ISSN: 1477-9234
Titre abrégé: Dalton Trans
Pays: England
ID NLM: 101176026
Informations de publication
Date de publication:
04 Aug 2021
04 Aug 2021
Historique:
pubmed:
10
7
2021
medline:
24
12
2021
entrez:
9
7
2021
Statut:
ppublish
Résumé
The organometallic H-cluster of the [FeFe]-hydrogenase consists of a [4Fe-4S] cubane bridged via a cysteinyl thiolate to a 2Fe subcluster ([2Fe]H) containing CO, CN-, and dithiomethylamine (DTMA) ligands. The H-cluster is synthesized by three dedicated maturation proteins: the radical SAM enzymes HydE and HydG synthesize the non-protein ligands, while the GTPase HydF serves as a scaffold for assembly of [2Fe]H prior to its delivery to the [FeFe]-hydrogenase containing the [4Fe-4S] cubane. HydG uses l-tyrosine as a substrate, cleaving it to produce p-cresol as well as the CO and CN- ligands to the H-cluster, although there is some question as to whether these are formed as free diatomics or as part of a [Fe(CO)2(CN)] synthon. Here we show that Clostridium acetobutylicum (C.a.) HydG catalyzes formation of multiple equivalents of free CO at rates comparable to those for CN- formation. Free CN- is also formed in excess molar equivalents over protein. A g = 8.9 EPR signal is observed for C.a. HydG reconstituted to load the 5th "dangler" iron of the auxiliary [4Fe-4S][FeCys] cluster and is assigned to this "dangler-loaded" cluster state. Free CO and CN- formation and the degree of activation of [FeFe]-hydrogenase all occur regardless of dangler loading, but are increased 10-35% in the dangler-loaded HydG; this indicates the dangler iron is not essential to this process but may affect relevant catalysis. During HydG turnover in the presence of myoglobin, the g = 8.9 signal remains unchanged, indicating that a [Fe(CO)2(CN)(Cys)] synthon is not formed at the dangler iron. Mutation of the only protein ligand to the dangler iron, H272, to alanine nearly completely abolishes both free CO formation and hydrogenase activation, however results show this is not due solely to the loss of the dangler iron. In experiments with wild type and H272A HydG, and with different degrees of dangler loading, we observe a consistent correlation between free CO/CN- formation and hydrogenase activation. Taken in full, our results point to free CO/CN-, but not an [Fe(CO)2(CN)(Cys)] synthon, as essential species in hydrogenase maturation.
Identifiants
pubmed: 34240096
doi: 10.1039/d1dt01359a
pmc: PMC9154046
mid: NIHMS1807888
doi:
Substances chimiques
Iron-Sulfur Proteins
0
Hydrogenase
EC 1.12.7.2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
10405-10422Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM054608
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM138592
Pays : United States
Organisme : NIGMS NIH HHS
ID : R29 GM054608
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM131889
Pays : United States
Références
Biochemistry. 2017 Jun 27;56(25):3234-3247
pubmed: 28525271
J Am Chem Soc. 2016 Feb 3;138(4):1146-9
pubmed: 26764535
Curr Opin Chem Biol. 2011 Apr;15(2):319-27
pubmed: 21393052
Biochemistry. 1999 Oct 5;38(40):12969-73
pubmed: 10529166
J Am Chem Soc. 2018 Apr 25;140(16):5516-5526
pubmed: 29595965
J Biol Chem. 2015 Feb 13;290(7):3987-94
pubmed: 25477518
Biochemistry. 2014 Jul 1;53(25):4090-104
pubmed: 24878200
Proc Natl Acad Sci U S A. 2008 Oct 21;105(42):16137-41
pubmed: 18852451
Nat Chem Biol. 2013 Oct;9(10):607-609
pubmed: 23934246
Proteins. 2017 Mar;85(3):435-444
pubmed: 27936493
Proc Natl Acad Sci U S A. 2009 Oct 13;106(41):17331-6
pubmed: 19805068
Chembiochem. 2015 Feb 9;16(3):397-402
pubmed: 25504963
FEBS Lett. 2011 Jan 3;585(1):225-30
pubmed: 21130763
Chem Rev. 2014 Apr 23;114(8):4149-74
pubmed: 24521136
Annu Rev Physiol. 1990;52:1-25
pubmed: 2184753
Angew Chem Int Ed Engl. 2010 Feb 22;49(9):1687-90
pubmed: 20108298
Science. 2016 May 13;352(6287):822-5
pubmed: 27174986
Nature. 2010 May 13;465(7295):248-51
pubmed: 20418861
Chem Sci. 2019 Dec 18;11(5):1241-1247
pubmed: 34123248
Proteins. 2013 Dec;81(12):2159-66
pubmed: 23996272
J Biol Inorg Chem. 2020 Aug;25(5):777-788
pubmed: 32661785
J Biol Inorg Chem. 2019 Sep;24(6):783-792
pubmed: 31493152
J Am Chem Soc. 2002 Feb 13;124(6):912-3
pubmed: 11829592
J Biol Chem. 2006 Jan 13;281(2):769-74
pubmed: 16278209
Int J Mol Sci. 2018 Oct 11;19(10):
pubmed: 30314343
Proc Natl Acad Sci U S A. 2016 Jan 5;113(1):104-9
pubmed: 26699472
J Am Chem Soc. 2002 Mar 27;124(12):3143-51
pubmed: 11902903
J Bacteriol. 2006 Mar;188(6):2163-72
pubmed: 16513746
J Am Chem Soc. 2010 Aug 25;132(33):11496-503
pubmed: 20684546
Science. 2014 Jan 24;343(6169):424-7
pubmed: 24458644
Proc Natl Acad Sci U S A. 2013 Apr 30;110(18):7188-92
pubmed: 23596207
Trends Biotechnol. 2014 Apr;32(4):170-6
pubmed: 24630475
Biochemistry. 2013 Dec 3;52(48):8696-707
pubmed: 24206022
Proc Natl Acad Sci U S A. 2015 Sep 15;112(37):11455-60
pubmed: 26324916
Proc Natl Acad Sci U S A. 2015 Feb 3;112(5):1362-7
pubmed: 25605932
J Biol Chem. 1989 Dec 15;264(35):20974-83
pubmed: 2556390
J Am Chem Soc. 2015 Feb 11;137(5):1809-16
pubmed: 25579778
Biochemistry. 2016 Jun 28;55(25):3514-27
pubmed: 27232385
J Biol Chem. 1990 Feb 25;265(6):3168-76
pubmed: 2303446
Dalton Trans. 2019 May 7;48(18):5978-5986
pubmed: 30632592
FEBS Lett. 2010 Oct 8;584(19):4197-202
pubmed: 20837009
Biochemistry. 1997 Sep 30;36(39):11811-20
pubmed: 9305972
J Inorg Biochem. 2011 Jun;105(6):784-92
pubmed: 21497576
FEBS Lett. 2010 Feb 5;584(3):638-42
pubmed: 20018187
J Biol Chem. 2008 Jul 4;283(27):18861-72
pubmed: 18400755
Nat Chem. 2017 Jan;9(1):88-95
pubmed: 27995927
Acc Chem Res. 2015 Aug 18;48(8):2380-7
pubmed: 26165393
J Am Chem Soc. 2002 Sep 25;124(38):11270-1
pubmed: 12236732
Curr Opin Biotechnol. 2010 Jun;21(3):244-51
pubmed: 20303737
Proc Natl Acad Sci U S A. 2019 Oct 15;116(42):20850-20855
pubmed: 31570604
Biochim Biophys Acta. 2015 Jun;1853(6):1350-69
pubmed: 25461840
Biochim Biophys Acta. 2000 Feb 9;1476(2):368-71
pubmed: 10669801
Proc Natl Acad Sci U S A. 2010 Jun 8;107(23):10448-53
pubmed: 20498089
Biochem J. 2017 Jan 15;474(2):215-230
pubmed: 28062838
Sci Rep. 2017 May 10;7(1):1714
pubmed: 28490758
Biochemistry. 1978 Jan 10;17(1):1-8
pubmed: 618535
Angew Chem Int Ed Engl. 2021 Feb 23;60(9):4666-4672
pubmed: 33935588
Biochemistry. 2017 Sep 12;56(36):4733-4734
pubmed: 28853860
Biochemistry. 2015 Mar 10;54(9):1807-18
pubmed: 25654171
FEBS Lett. 2008 Jun 25;582(15):2183-7
pubmed: 18501709
Chem Rev. 2014 Apr 23;114(8):4081-148
pubmed: 24655035
J Biol Chem. 1993 Jul 15;268(20):14732-42
pubmed: 8325851
Science. 2013 Oct 25;342(6157):472-5
pubmed: 24159045
Structure. 2020 Sep 1;28(9):1071-1081.e3
pubmed: 32649857
Biochem Biophys Res Commun. 2000 Mar 16;269(2):451-6
pubmed: 10708574
J Biol Chem. 2011 Nov 25;286(47):40614-23
pubmed: 21930709
J Am Chem Soc. 2021 Jan 13;143(1):335-348
pubmed: 33372786
Nat Chem Biol. 2017 Jul;13(7):779-784
pubmed: 28553946
FEBS Lett. 2009 Feb 4;583(3):506-11
pubmed: 19166853
Nature. 2013 Jul 4;499(7456):66-69
pubmed: 23803769
J Biol Chem. 1984 Dec 10;259(23):14463-71
pubmed: 6094558
Dalton Trans. 2018 Jul 17;47(28):9521-9535
pubmed: 29964288
PLoS One. 2011;6(5):e20346
pubmed: 21673792
J Am Chem Soc. 2020 Jun 17;142(24):10841-10848
pubmed: 32434327
PLoS One. 2012;7(9):e45850
pubmed: 23049878
Structure. 2011 Aug 10;19(8):1038-52
pubmed: 21827941
J Biol Chem. 2020 Aug 14;295(33):11891-11901
pubmed: 32620553
Eur J Biochem. 1987 Jan 2;162(1):31-6
pubmed: 3028789
J Biol Chem. 1999 Feb 5;274(6):3331-7
pubmed: 9920874
J Mol Biol. 1993 Nov 5;234(1):140-55
pubmed: 8230194
Annu Rev Plant Biol. 2007;58:71-91
pubmed: 17150028
J Am Chem Soc. 2013 May 8;135(18):6921-9
pubmed: 23578101
J Am Chem Soc. 2015 Oct 7;137(39):12580-7
pubmed: 26352172
J Am Chem Soc. 2014 Sep 24;136(38):13086-9
pubmed: 25099480
J Am Chem Soc. 2010 Jul 14;132(27):9247-9
pubmed: 20565074
Sustain Energy Fuels. 2018 Apr 1;2(4):724-750
pubmed: 31497651
Science. 1998 Dec 4;282(5395):1853-8
pubmed: 9836629
J Biol Chem. 1990 May 25;265(15):8533-41
pubmed: 2160461
J Biol Chem. 2012 Oct 19;287(43):36544-55
pubmed: 22932901
Nat Chem. 2016 May;8(5):491-500
pubmed: 27102684
J Biol Chem. 2004 Jun 11;279(24):25711-20
pubmed: 15082711
Nat Protoc. 2017 Feb;12(2):255-278
pubmed: 28079879
Photosynth Res. 2015 Sep;125(3):383-93
pubmed: 26022106
J Am Chem Soc. 2019 Oct 9;141(40):16117-16124
pubmed: 31509404
Biochemistry. 2009 Jul 7;48(26):6240-8
pubmed: 19435321
J Biol Inorg Chem. 2007 May;12(4):443-7
pubmed: 17372774
Biochemistry. 2004 Aug 10;43(31):10203-11
pubmed: 15287748
Curr Opin Biotechnol. 2012 Jun;23(3):346-51
pubmed: 22104720
J Biol Chem. 1985 Sep 15;260(20):11160-73
pubmed: 2993304
Biochemistry. 2000 Jun 27;39(25):7455-60
pubmed: 10858294
Nat Chem. 2018 May;10(5):555-560
pubmed: 29632334
Methods Enzymol. 2018;606:269-318
pubmed: 30097096
Photosynth Res. 2015 Mar;123(3):241-53
pubmed: 24671643