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1. Next generation single-molecule techniques: Imaging, labeling, and manipulation in vitro and in cellulo

   https://doi.org/10.1016/j.molcel.2021.12.019  

   Ha, Taekjip ; Kaiser, Christian ; Myong, Sua ; Wu, Bin ; Xiao, Jie ( January 2022 , Molecular Cell)
2. Access Balancing in Storage Systems by Labeling Partial Steiner Systems

   https://doi.org/10.1109/ISIT44484.2020.9174154  

   Chee, Y. M. ; Colbourn, C. J. ; Dau, H. ; Gabrys, R. ; Ling, A. C. ; Lusi, D. ; Milenkovic, O. ( June 2020 , 2020 IEEE International Symposium on Information Theory (ISIT))

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3. Spin-Labeling Insights into How Chemical Fixation Impacts Glycan Organization on Cells

   https://doi.org/10.1007/s00723-023-01624-w  

   Jaiswal, Mohit ; Tran, Trang T. ; Guo, Jiatong ; Zhou, Mingwei ; Kundu, Sayan ; Guo, Zhongwu ; Fanucci, Gail E. ( October 2023 , Applied Magnetic Resonance)

   Zhongyu Yang (Ed.)

   As new methods to interrogate glycan organization on cells develop, it is important to have a molecular level understanding of how chemical fixation can impact results and interpretations. Site-directed spin labeling technologies are well suited to study how the spin label mobility is impacted by local environmental conditions, such as those imposed by cross-linking effects of paraformaldehyde cell fixation methods. Here, we utilize three different azide-containing sugars for metabolic glycan engineering with HeLa cells to incorporate azido glycans that are modified with a DBCO-based nitroxide moiety via click reaction. Continuous wave X-band electron paramagnetic resonance spectroscopy is employed to characterize how the chronological sequence of chemical fixation and spin labeling impacts the local mobility and accessibility of the nitroxide-labeled glycans in the glycocalyx of HeLa cells. Results demonstrate that chemical fixation with paraformaldehyde can alter local glycan mobility and care should be taken in the analysis of data in any study where chemical fixation and cellular labeling occur. 

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4. Isotopic Labeling of Formate Dehydrogenase Perturbs the Protein Dynamics

   https://doi.org/10.1021/acs.jpcb.9b08426  

   Ranasinghe, Chethya ; Pagano, Philip L ; Sapienza, Paul J. ; Lee, Andrew L. ; Kohen, Amnon ; Cheatum, Christopher M. ( November 2019 , The Journal of Physical Chemistry B)
5. Sortase-mediated segmental labeling: A method for segmental assignment of intrinsically disordered regions in proteins

   https://doi.org/10.1371/journal.pone.0258531  

   Boyko, Kristina V. ; Rosenkranz, Erin A. ; Smith, Derrick M. ; Miears, Heather L. ; Oueld es cheikh, Melissa ; Lund, Micah Z. ; Young, Jeffery C. ; Reardon, Patrick N. ; Okon, Mark ; Smirnov, Serge L. ; et al ( October 2021 , PLOS ONE)

   van der Wel, Patrick (Ed.)

   A significant number of proteins possess sizable intrinsically disordered regions (IDRs). Due to the dynamic nature of IDRs, NMR spectroscopy is often the tool of choice for characterizing these segments. However, the application of NMR to IDRs is often hindered by their instability, spectral overlap and resonance assignment difficulties. Notably, these challenges increase considerably with the size of the IDR. In response to these issues, here we report the use of sortase-mediated ligation (SML) for segmental isotopic labeling of IDR-containing samples. Specifically, we have developed a ligation strategy involving a key segment of the large IDR and adjacent folded headpiece domain comprising the C-terminus of A . thaliana villin 4 (AtVLN4). This procedure significantly reduces the complexity of NMR spectra and enables group identification of signals arising from the labeled IDR fragment, a process we refer to as segmental assignment . The validity of our segmental assignment approach is corroborated by backbone residue-specific assignment of the IDR using a minimal set of standard heteronuclear NMR methods. Using segmental assignment, we further demonstrate that the IDR region adjacent to the headpiece exhibits nonuniform spectral alterations in response to temperature. Subsequent residue-specific characterization revealed two segments within the IDR that responded to temperature in markedly different ways. Overall, this study represents an important step toward the selective labeling and probing of target segments within much larger IDR contexts. Additionally, the approach described offers significant savings in NMR recording time, a valuable advantage for the study of unstable IDRs, their binding interfaces, and functional mechanisms. 

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