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Investigating mycobacteria cell wall formation using FDAA HADA.
Investigating mycobacteria cell wall formation using fluorescent D-amino acid HADA. Image showing FDAA HADA incorporation in log-phase growth of WT, delta L,D-transpeptidases (LDT) and delta L,D-transpeptidases (LDT) comp cells. Scale = 5 μm. Baranowski et al show FDAAs are incorporated asymmetrically by L,D-transpeptidases. For more information on this work and to request full protocols read Baranowski et al (2018) Maturing Mycobacterium smegmatis peptidoglycan requires non-canonical crosslinks to maintain shape. Elife 16 e37516 PMID: 30324906
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Using FDAAs HADA and RADA to study peptidoglycan enzyme and substrate distribution, and the role of polar cell growth and peptidoglycan segregation over time.
Image shows peptidoglycan segregation over time. Left to right - 2 mins HADA pulse (cyan), 45 min outgrowth, followed by 2 min RADA (Catalog # 6649) chase (magenta) in WT Msm cells. New cell wall (magenta), oldest cell wall (cyan). Scale = 5 μm. For more information on this work and to request full protocols please read García-Heredia et al (2018) Peptidoglycan precursor synthesis along the sidewall of pole-growing mycobacteria. Elife 10 e37243 PMID: 30198841
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