{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T07:58:23Z","timestamp":1781855903947,"version":"3.54.5"},"posted":{"date-parts":[[2026,5,20]]},"group-title":"Microbiology","reference-count":52,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T00:00:00Z","timestamp":1779235200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,20]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  We present an ultrahigh-field magic-angle spinning (MAS) solid-state NMR (ssNMR) study to characterize intact nontuberculous mycobacteria (NTM) at the molecular level. Hydrated and dried whole-cell\n                  <jats:italic>Mycobacterium abscessus<\/jats:italic>\n                  samples were investigated by combining conventional high-field ssNMR at 750 MHz with ultrahigh-field ssNMR at 1.2 GHz and ultrafast MAS at 100 kHz. To improve sensitivity and enable multidimensional experiments, 13C\/15N isotope labeling was performed after growth in synthetic cystic fibrosis medium (SCFM). We utilized 1D 13C and multidimensional 1H-13C and 13C-13C ssNMR experiments to characterize the chemical composition, dynamics, and structural organization of the\n                  <jats:italic>M. abscessus<\/jats:italic>\n                  cell envelope. The isotope-labeling efficiency was found to be non-uniform across different molecular classes, with high incorporation into polysaccharides and lower incorporation into lipid and peptide-associated signals. INEPT- and CP-based experiments selectively probed flexible and rigid fractions of the samples, revealing substantial differences in linewidth, dynamics, and sensitivity between hydrated and dried preparations. Conventional 750 MHz experiments provided high-resolution multidimensional spectra and enabled identification of distinct chemical environments associated with peptidoglycan, arabinogalactan, mycolic acids, lipids, and peptide-associated components. Ultrahigh-field ssNMR at 1.2 GHz combined with ultrafast MAS and 1H detection substantially improved spectral resolution and sensitivity in particular per mg of sample amount, allowing detection of weak and previously unresolved resonances, including polysaccharide and possible nucleic-acid-associated signals. Together, these results demonstrate that ultra-high-field and ultrafast-MAS ssNMR enables detailed characterization of intact NTM cell envelopes under near-native conditions and provides a framework for future molecular investigations of antimicrobial interactions.\n                <\/jats:p>","DOI":"10.64898\/2026.05.19.726312","type":"posted-content","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T04:30:17Z","timestamp":1779337817000},"source":"Crossref","is-referenced-by-count":1,"title":["Structural Insights into Native Intact\n                  <i>Mycobacterium abscessus<\/i>\n                  by Conventional and Ultrahigh-field solid-state NMR at 1.2 GHz"],"prefix":"10.64898","author":[{"given":"Chang-Hyeock","family":"Byeon","sequence":"first","affiliation":[{"name":"Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, 15261, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Hao","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, University of Pittsburgh, Pittsburgh, 15261, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdulkadir","family":"Tunc","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, 15261, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. Trent","family":"Franks","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Campus Chemical Instrument Center NMR Facility, The Ohio State University, Columbus, OH, 43210, United States"},{"name":"National Gateway Ultrahigh Field NMR Center, The Ohio State University, Columbus, OH, 43210, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William H.","family":"DePas","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, University of Pittsburgh, Pittsburgh, 15261, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00dcmit","family":"Akbey","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, 15261, United 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