BIOMAT Database Logo BIOMAT Database Logo

Quick and Spontaneous Transformation between [3Fe-4S] and [4Fe-4S] Iron-Sulfur Clusters in the tRNA-Thiolation Enzyme TtuA. 2023

Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
Graduate School of Life Science, Hokkaido University, Kita 8, Nishi 5, Kita-ku, Sapporo 060-0810, Japan.

Iron-sulfur (Fe-S) clusters are essential cofactors for enzyme activity. These Fe-S clusters are present in structurally diverse forms, including [4Fe-4S] and [3Fe-4S]. Type-identification of the Fe-S cluster is indispensable in understanding the catalytic mechanism of enzymes. However, identifying [4Fe-4S] and [3Fe-4S] clusters in particular is challenging because of their rapid transformation in response to oxidation-reduction events. In this study, we focused on the relationship between the Fe-S cluster type and the catalytic activity of a tRNA-thiolation enzyme (TtuA). We reconstituted [4Fe-4S]-TtuA, prepared [3Fe-4S]-TtuA by oxidizing [4Fe-4S]-TtuA under strictly anaerobic conditions, and then observed changes in the Fe-S clusters in the samples and the enzymatic activity in the time-course experiments. Electron paramagnetic resonance analysis revealed that [3Fe-4S]-TtuA spontaneously transforms into [4Fe-4S]-TtuA in minutes to one hour without an additional free Fe source in the solution. Although the TtuA immediately after oxidation of [4Fe-4S]-TtuA was inactive [3Fe-4S]-TtuA, its activity recovered to a significant level compared to [4Fe-4S]-TtuA after one hour, corresponding to an increase of [4Fe-4S]-TtuA in the solution. Our findings reveal that [3Fe-4S]-TtuA is highly inactive and unstable. Moreover, time-course analysis of structural changes and activity under strictly anaerobic conditions further unraveled the Fe-S cluster type used by the tRNA-thiolation enzyme.

UI MeSH Term Description Entries
D007501 Iron A metallic element with atomic symbol Fe, atomic number 26, and atomic weight 55.85. It is an essential constituent of HEMOGLOBINS; CYTOCHROMES; and IRON-BINDING PROTEINS. It plays a role in cellular redox reactions and in the transport of OXYGEN. Iron-56,Iron 56
D007506 Iron-Sulfur Proteins A group of proteins possessing only the iron-sulfur complex as the prosthetic group. These proteins participate in all major pathways of electron transport: photosynthesis, respiration, hydroxylation and bacterial hydrogen and nitrogen fixation. Iron-Sulfur Protein,Iron Sulfur Proteins,Iron Sulfur Protein,Protein, Iron-Sulfur,Proteins, Iron Sulfur,Proteins, Iron-Sulfur,Sulfur Proteins, Iron
D010084 Oxidation-Reduction A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471). Redox,Oxidation Reduction
D004578 Electron Spin Resonance Spectroscopy A technique applicable to the wide variety of substances which exhibit paramagnetism because of the magnetic moments of unpaired electrons. The spectra are useful for detection and identification, for determination of electron structure, for study of interactions between molecules, and for measurement of nuclear spins and moments. (From McGraw-Hill Encyclopedia of Science and Technology, 7th edition) Electron nuclear double resonance (ENDOR) spectroscopy is a variant of the technique which can give enhanced resolution. Electron spin resonance analysis can now be used in vivo, including imaging applications such as MAGNETIC RESONANCE IMAGING. ENDOR,Electron Nuclear Double Resonance,Electron Paramagnetic Resonance,Paramagnetic Resonance,Electron Spin Resonance,Paramagnetic Resonance, Electron,Resonance, Electron Paramagnetic,Resonance, Electron Spin,Resonance, Paramagnetic
D012343 RNA, Transfer The small RNA molecules, 73-80 nucleotides long, that function during translation (TRANSLATION, GENETIC) to align AMINO ACIDS at the RIBOSOMES in a sequence determined by the mRNA (RNA, MESSENGER). There are about 30 different transfer RNAs. Each recognizes a specific CODON set on the mRNA through its own ANTICODON and as aminoacyl tRNAs (RNA, TRANSFER, AMINO ACYL), each carries a specific amino acid to the ribosome to add to the elongating peptide chains. Suppressor Transfer RNA,Transfer RNA,tRNA,RNA, Transfer, Suppressor,Transfer RNA, Suppressor,RNA, Suppressor Transfer
D013455 Sulfur An element that is a member of the chalcogen family. It has an atomic symbol S, atomic number 16, and atomic weight [32.059; 32.076]. It is found in the amino acids cysteine and methionine. Sulfur-16,Sulfur 16

Related Publications

Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
Structure and electrochemistry of proteins harboring iron-sulfur clusters of different nuclearities. Part II. [4Fe-4S] and [3Fe-4S] iron-sulfur proteins.
June 2018, Journal of structural biology,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
Structure and electrochemistry of proteins harboring iron-sulfur clusters of different nuclearities. Part III. [4Fe-4S], [3Fe-4S] and [2Fe-2S] iron-sulfur proteins.
June 2018, Journal of structural biology,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
Nonredox thiolation in tRNA occurring via sulfur activation by a [4Fe-4S] cluster.
July 2017, Proceedings of the National Academy of Sciences of the United States of America,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
Iron-sulfur stoichiometry and structure of iron-sulfur clusters in three-iron proteins: evidence for [3Fe-4S] clusters.
January 1983, Proceedings of the National Academy of Sciences of the United States of America,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
[4Fe-4S]-cluster-depleted Azotobacter vinelandii ferredoxin I: a new 3Fe iron-sulfur protein.
September 1985, Proceedings of the National Academy of Sciences of the United States of America,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
MiaB, a bifunctional radical-S-adenosylmethionine enzyme involved in the thiolation and methylation of tRNA, contains two essential [4Fe-4S] clusters.
May 2007, Biochemistry,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
HdrC2 from Acidithiobacillus ferrooxidans owns two iron-sulfur binding motifs but binds only one variable cluster between [4Fe-4S] and [3Fe-4S].
January 2013, Current microbiology,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
A [3Fe-4S] cluster is required for tRNA thiolation in archaea and eukaryotes.
November 2016, Proceedings of the National Academy of Sciences of the United States of America,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
Formation of [4Fe-4S] clusters in the mitochondrial iron-sulfur cluster assembly machinery.
November 2014, Journal of the American Chemical Society,
Masato Ishizaka, and Minghao Chen, and Shun Narai, and Yoshikazu Tanaka, and Toyoyuki Ose, and Masaki Horitani, and Min Yao
A pathway for assembling [4Fe-4S]2+ clusters in mitochondrial iron-sulfur protein biogenesis.
June 2020, The FEBS journal,
Copied contents to your clipboard!