Engineering of an alternative electron transfer path in photosystem II.
The initial steps of oxygenic photosynthetic electron transfer occur within photosystem II, an intricate pigment/protein transmembrane complex. Light-driven electron transfer occurs within a multistep pathway that is efficiently insulated from competing electron transfer pathways. The heart of the electron transfer system, composed of six linearly coupled redox active cofactors that enable electron transfer from water to the secondary quinone acceptor Q(B), is mainly embedded within two proteins called D1 and D2. We have identified a site in silico, poised in the vicinity of the Q(A) intermediate quinone acceptor, which could serve as a potential binding site for redox active proteins. Here we show that modification of Lysine 238 of the D1 protein to glutamic acid (Glu) in the cyanobacterium Synechocystis sp. PCC 6803, results in a strain that grows photautotrophically. The Glu thylakoid membranes are able to perform light-dependent reduction of exogenous cytochrome c with water as the electron donor. Cytochrome c photoreduction by the Glu mutant was also shown to significantly protect the D1 protein from photodamage when isolated thylakoid membranes were illuminated. We have therefore engineered a novel electron transfer pathway from water to a soluble protein electron carrier without harming the normal function of photosystem II.
Larom
Shirley
S
Faculty of Biology and Schulich Faculty of Chemistry, Technion-Israel Institute of Technology, Haifa 32000, Israel.
Salama
Faris
F
Schuster
Gadi
G
Adir
Noam
N
D001665:Binding Sites; D019295:Computational Biology; D045304:Cytochromes c / Q000378:metabolism; D004579:Electron Transport; D008958:Models, Molecular; D009154:Mutation; D045332:Photosystem II Protein Complex / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D015202:Protein Engineering; D020836:Protein Structure, Quaternary; D046939:Synechocystis / Q000201:enzymology*; D020524:Thylakoids / Q000201:enzymology
D045332:Photosystem II Protein Complex; D045304:Cytochromes c
Nucleic Acids Res. 2007 Jul;35(Web Server issue):W522-5
17488841
Photosynth Res. 2003;76(1-3):343-70
16228592
Biochim Biophys Acta. 1999 Nov 10;1413(3):147-58
10556627
Trends Biotechnol. 2007 Oct;25(10):437-40
17870196
J Biol Chem. 2007 Apr 27;282(17):12492-502
17327225
Biotechnol Prog. 2008 Jul-Aug;24(4):815-20
18335954
Plant Mol Biol. 2000 Mar;42(4):635-45
10809009
Biochim Biophys Acta. 2007 Jun;1767(6):414-21
17207454
Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10037-41
11517324
Photochem Photobiol Sci. 2005 Dec;4(12):957-70
16307108
Annu Rev Plant Biol. 2002;53:449-75
12221984
Biochemistry. 1988 Apr 5;27(7):2450-61
2838074
Cell Mol Life Sci. 2009 Dec;66(23):3697-710
19644734
Photosynth Res. 2008 Oct-Dec;98(1-3):53-80
18830685
J Ind Microbiol Biotechnol. 2009 Feb;36(2):269-74
18982369
Adv Protein Chem. 2003;63:71-109
12629967
Plant Cell. 1991 Apr;3(4):383-95
1840918
Nature. 2005 Dec 15;438(7070):1040-4
16355230
Photochem Photobiol. 2004 Feb;79(2):152-62
15068028
Nat Rev Mol Cell Biol. 2004 Dec;5(12):971-82
15573135
Nat Struct Mol Biol. 2009 Mar;16(3):334-42
19219048
Eur J Biochem. 1988 Nov 1;177(2):403-10
3191923
Science. 2004 Mar 19;303(5665):1831-8
14764885
Chem Soc Rev. 2009 Apr;38(4):892-901
19421569
Photosynth Res. 2008 Oct-Dec;98(1-3):589-608
18937045
Eur J Biochem. 1996 May 1;237(3):519-31
8647094
Chem Soc Rev. 2009 Jan;38(1):185-96
19088973
Photosynth Res. 2006 Jan;87(1):25-40
16416050
EMBO J. 1988 Jan;7(1):1-6
16453819
J Mol Biol. 2002 May 31;319(2):501-15
12051924
Q Rev Biophys. 2003 Aug;36(3):341-72
15029828
FEBS Lett. 2003 Nov 27;555(1):45-50
14630317
Annu Rev Plant Biol. 2009;60:165-82
19014348
Photosynth Res. 2005;85(1):101-14
15977062
Curr Opin Biotechnol. 2008 Jun;19(3):235-40
18539450
Results Probl Cell Differ. 2008;45:33-72
18066506
Photosynth Res. 2008 Oct-Dec;98(1-3):643-55
18853275
Biochemistry. 1996 Feb 27;35(8):2535-47
8611557
J Mol Biol. 2007 Aug 10;371(2):396-409
17570397
Photosynth Res. 2008 Oct-Dec;98(1-3):489-501
18618287
Annu Rev Plant Biol. 2006;57:521-65
16669773
Curr Opin Biotechnol. 2009 Jun;20(3):257-63
19540103
Photosynth Res. 2008 Oct-Dec;98(1-3):687-700
18777102
Biochemistry. 1998 Dec 15;37(50):17339-44
9860848
Recent Pat Biotechnol. 2008;2(1):25-34
19075850