Analyzing the structure of macromolecules in their native cellular environment using hydroxyl radical footprinting

The Analyst ◽  
2018 ◽  
Vol 143 (4) ◽  
pp. 798-807 ◽  
Author(s):  
Emily E. Chea ◽  
Lisa M. Jones
Keyword(s):  
Nucleic Acids ◽  
Hydroxyl Radical ◽  
Live Cells ◽  
Cellular Environment ◽  
Hydroxyl Radical Footprinting

Hydroxyl radical footprinting (HRF) has been successfully used to study the structure of both nucleic acids and proteins in live cells.

scholarly journals POT1-TPP1 differentially regulates telomerase via POT1 His266 and as a function of single-stranded telomere DNA length

2019 ◽  
Vol 116 (47) ◽  
pp. 23527-23533 ◽  
Author(s):  
Mengyuan Xu ◽  
Janna Kiselar ◽  
Tawna L. Whited ◽  
Wilnelly Hernandez-Sanchez ◽  
Derek J. Taylor
Keyword(s):  
Conformational Changes ◽  
Protein Interactions ◽  
Environmental Changes ◽  
Lymphocytic Leukemia ◽  
Protein Protein Interactions ◽  
Cellular Environment ◽  
Multiple Protein ◽  
Linear Chromosomes ◽  
Hydroxyl Radical Footprinting ◽  
Regulate Telomere Length

Telomeres cap the ends of linear chromosomes and terminate in a single-stranded DNA (ssDNA) overhang recognized by POT1-TPP1 heterodimers to help regulate telomere length homeostasis. Here hydroxyl radical footprinting coupled with mass spectrometry was employed to probe protein–protein interactions and conformational changes involved in the assembly of telomere ssDNA substrates of differing lengths bound by POT1-TPP1 heterodimers. Our data identified environmental changes surrounding residue histidine 266 of POT1 that were dependent on telomere ssDNA substrate length. We further determined that the chronic lymphocytic leukemia-associated H266L substitution significantly reduced POT1-TPP1 binding to short ssDNA substrates; however, it only moderately impaired the heterodimer binding to long ssDNA substrates containing multiple protein binding sites. Additionally, we identified a telomerase inhibitory role when several native POT1-TPP1 proteins coat physiologically relevant lengths of telomere ssDNA. This POT1-TPP1 complex-mediated inhibition of telomerase is abrogated in the context of the POT1 H266L mutation, which leads to telomere overextension in a malignant cellular environment.

scholarly journals Mapping Functionally Important Motifs SPF and GGQ of the Decoding Release Factor RF2 to theEscherichia coliRibosome by Hydroxyl Radical Footprinting

2002 ◽  
Vol 278 (17) ◽  
pp. 15095-15104 ◽  
Author(s):  
Debbie-Jane G. Scarlett ◽  
Kim K. McCaughan ◽  
Daniel N. Wilson ◽  
Warren P. Tate
Keyword(s):  
Hydroxyl Radical ◽  
Release Factor ◽  
Hydroxyl Radical Footprinting

scholarly journals Structural interpretation of DNA–protein hydroxyl-radical footprinting experiments with high resolution using HYDROID

2018 ◽  
Vol 13 (11) ◽  
pp. 2535-2556 ◽  
Author(s):  
Alexey K. Shaytan ◽  
Hua Xiao ◽  
Grigoriy A. Armeev ◽  
Daria A. Gaykalova ◽  
Galina A. Komarova ◽  
...  
Keyword(s):  
High Resolution ◽  
Hydroxyl Radical ◽  
Structural Interpretation ◽  
Hydroxyl Radical Footprinting
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