Structural and Biophysical Properties of a [4Fe-4S] Ferredoxin-Like Protein from Synechocystis sp. PCC 6803 with a Unique Two Domain Structure

Kisgeropoulos, Effie; Bharadwaj, Vivek S.; Ledinina, Anastasia; Lubner, Carolyn E.; Mulder, David W.; Smolinski, Sharon L.; Boehm, Marko; Gutekunst, Kirstin; King, Paul W.; Svedruzic, Drazenka

doi:10.1016/j.jinorgbio.2023.112428

Structural and Biophysical Properties of a [4Fe-4S] Ferredoxin-Like Protein from Synechocystis sp. PCC 6803 with a Unique Two Domain Structure

Journal Article · Sat Nov 18 23:00:00 EST 2023 · Journal of Inorganic Biochemistry

DOI:https://doi.org/10.1016/j.jinorgbio.2023.112428· OSTI ID:2280799

; ; Ledinina, Anastasia; ; Mulder, David W.; Smolinski, Sharon L.; Boehm, Marko; Gutekunst, Kirstin; ;

Electron carrier proteins (ECPs), binding iron-sulfur clusters, are vital components within the intricate network of metabolic and photosynthetic reactions. They play a crucial role in the distribution of reducing equivalents. In Synechocystis sp. PCC 6803, the ECP network includes at least nine ferredoxins. Previous research, including global expression analyses and protein binding studies, has offered initial insights into the functional roles of individual ferredoxins within this network. This study primarily focuses on Ferredoxin 9 (slr2059). Through sequence analysis and computational modeling, Ferredoxin 9 emerges as a unique ECP with a distinctive two-domain architecture. It consists of a C-terminal iron-sulfur binding domain and an N-terminal domain with homology to Nil-domain proteins, connected by a structurally rigid 4-amino acid linker. Notably, in contrast to canonical [2Fe-2S] ferredoxins exemplified by PetF (ssl0020), which feature highly acidic surfaces facilitating electron transfer with photosystem I reaction centers, models of Ferredoxin 9 reveal a more neutral to basic protein surface. Using a combination of electron paramagnetic resonance spectroscopy and square-wave voltammetry on heterologously produced Ferredoxin 9, this study demonstrates that the protein coordinates 2x[4Fe-4S]2+/1+ redox-active and magnetically interacting clusters, with measured redox potentials of -420 +/- 9 mV and -516 +/- 10 mV vs SHE. A more in-depth analysis of Fdx9's unique structure and protein sequence suggests that this type of Nil-2[4Fe-4S] multi-domain ferredoxin is well conserved in cyanobacteria, bearing structural similarities to proteins involved in homocysteine synthesis in methanogens.

Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division

DOE Contract Number:: AC36-08GO28308

OSTI ID:: 2280799

Report Number(s):: NREL/JA-2700-87453; MainId:88228; UUID:23788bd9-dd2c-496b-8503-c1b164bfe8cc; MainAdminID:71491

Journal Information:: Journal of Inorganic Biochemistry, Journal Name: Journal of Inorganic Biochemistry Vol. 251

Country of Publication:: United States

Language:: English

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INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
cyanobacteria
electron carrier proteins
metabolism
metalloproteins
oxidative stress
photosynthesis
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Structural and Biophysical Properties of a [4Fe-4S] Ferredoxin-Like Protein from Synechocystis sp. PCC 6803 with a Unique Two Domain Structure

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