scholarly journals Molecular Mechanism of ISC Iron–Sulfur Cluster Biogenesis Revealed by High-Resolution Native Mass Spectrometry

2020 ◽  
Vol 142 (13) ◽  
pp. 6018-6029 ◽  
Author(s):  
Cheng-Wei Lin ◽  
Jacob W. McCabe ◽  
David H. Russell ◽  
David P. Barondeau
Keyword(s):  
Mass Spectrometry ◽  
High Resolution ◽  
Molecular Mechanism ◽  
Native Mass Spectrometry ◽  
Iron Sulfur Cluster ◽  
Sulfur Cluster ◽  
Iron Sulfur

scholarly journals Sensing mechanisms of iron-sulfur cluster regulatory proteins elucidated using native mass spectrometry

2021 ◽  
Author(s):  
Jason C Crack ◽  
Elizabeth Gray ◽  
Nick Le Brun
Keyword(s):  
Mass Spectrometry ◽  
Native Mass Spectrometry ◽  
Regulatory Proteins ◽  
Environmental Cues ◽  
Iron Sulfur Cluster ◽  
Sulfur Cluster ◽  
Iron Sulfur ◽  
Sense And Respond

The ability to sense and respond to various key environmental cues is important for the survival and adaptability of many bacteria, including pathogens. The particular sensitivity of iron-sulfur (Fe-S) clusters...

scholarly journals An Iron-Sulfur Cluster in the C-terminal Domain of the p58 Subunit of Human DNA Primase

2007 ◽  
Vol 282 (46) ◽  
pp. 33444-33451 ◽  
Author(s):  
Brian E. Weiner ◽  
Hao Huang ◽  
Brian M. Dattilo ◽  
Mark J. Nilges ◽  
Ellen Fanning ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Dna Replication ◽  
Enzymatic Activity ◽  
Limited Proteolysis ◽  
High Potential ◽  
Multiple Sequence ◽  
Iron Sulfur Cluster ◽  
Sulfur Cluster ◽  
Dna Primase ◽  
Iron Sulfur

DNA primase synthesizes short RNA primers that are required to initiate DNA synthesis on the parental template strands during DNA replication. Eukaryotic primase contains two subunits, p48 and p58, and is normally tightly associated with DNA polymerase α. Despite the fundamental importance of primase in DNA replication, structural data on eukaryotic DNA primase are lacking. The p48/p58 dimer was subjected to limited proteolysis, which produced two stable structural domains: one containing the bulk of p48 and the other corresponding to the C-terminal fragment of p58. These domains were identified by mass spectrometry and N-terminal sequencing. The C-terminal p58 domain (p58C) was expressed, purified, and characterized. CD and NMR spectroscopy experiments demonstrated that p58C forms a well folded structure. The protein has a distinctive brownish color, and evidence from inductively coupled plasma mass spectrometry, UV-visible spectrophotometry, and EPR spectroscopy revealed characteristics consistent with the presence of a [4Fe-4S] high potential iron protein cluster. Four putative cysteine ligands were identified using a multiple sequence alignment, and substitution of just one was sufficient to cause loss of the iron-sulfur cluster and a reduction in primase enzymatic activity relative to the wild-type protein. The discovery of an iron-sulfur cluster in DNA primase that contributes to enzymatic activity provides the first suggestion that the DNA replication machinery may have redox-sensitive activities. Our results offer new horizons in which to investigate the function of high potential [4Fe-4S] clusters in DNA-processing machinery.

scholarly journals Iron-sulfur clusters are involved in post-translational arginylation

2021 ◽  
Author(s):  
Verna Van ◽  
Janae B. Brown ◽  
Hannah Rosenbach ◽  
Ijaz Mohamed ◽  
Nna-Emeka Ejimogu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Molecular Mechanism ◽  
Half Life ◽  
Regulatory Control ◽  
Post Translational Modification ◽  
Iron Sulfur Cluster ◽  
Iron Sulfur Clusters ◽  
Sulfur Cluster ◽  
Iron Sulfur ◽  
Oxygen Sensitive

Eukaryotic arginylation is an essential post-translational modification that both modulates protein stability and regulates protein half-life through the N-degron pathway. Arginylation is catalyzed by a family of enzymes known as the arginyl-tRNA transferases (ATE1s), which are conserved across the eukaryotic domain. Despite its conservation and importance, little is known regarding the structure, mechanism, and regulation of ATE1s. In this work, we have discovered that ATE1s bind a previously unknown iron-sulfur cluster that is conserved across evolution. We have extensively characterized the nature of this iron-sulfur cluster, and we show that the presence of the iron-sulfur cluster is linked to alterations in arginylation efficacy. Finally, we demonstrate that the ATE1 iron-sulfur cluster is oxygen sensitive, which could be a molecular mechanism of the N-degron pathway to sense oxidative stress. Thus, our data provide the framework of a cluster-based paradigm of ATE1 regulatory control.

Observation of the Iron-Sulfur Cluster inEscherichiacoliBiotin Synthase by Nanoflow Electrospray Mass Spectrometry

2001 ◽  
Vol 73 (17) ◽  
pp. 4154-4161 ◽  
Author(s):  
H. Hernández ◽  
K. S. Hewitson ◽  
P. Roach ◽  
N. M. Shaw ◽  
J. E. Baldwin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Mass Spectrometry ◽  
Iron Sulfur Cluster ◽  
Sulfur Cluster ◽  
Iron Sulfur
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