On-target labeling of intracellular metabolites combined with chemical mapping of individual hyphae revealing cytoplasmic relocation of isotopologues

Analytical Chemistry
Jie-Bi HuPawel L Urban

Abstract

A microscale analytical platform integrating microbial cell culture, isotopic labeling, along with visual and mass spectrometric imaging with single-cell resolution has been developed and applied in the monitoring of cellular metabolism in fungal mycelium. The method implements open chips with a two-dimensional surface pattern composed of hydrophobic and hydrophilic zones. Two hydrophilic islands are used as medium reservoirs, while the hydrophobic area constitutes the support for the growing aerial hyphae, which do not have direct contact with the medium. The first island, containing (12)C(6)-glucose medium, was initially inoculated with the mycelium (Neurospora crassa), and following the initial incubation period, the hyphae progressed toward the second medium island, containing an isotopically labeled substrate ((13)C(6)-glucose). The (13)C atoms were gradually incorporated into cellular metabolites, which was revealed by MALDI-MS. The fate of the chitin-biosynthesis precursor, uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), was monitored by recording mass spectra with characteristic isotopic patterns, which indicated the presence of various (12)C/(13)C isotopologues. The method enabled mapping the (13)C-labeled UDP-Gl...Continue Reading

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Dec 19, 2012·Current Opinion in Biotechnology·Stanislav S RubakhinJonathan V Sweedler
Sep 22, 2015·Analytica Chimica Acta·Jing WuJin-Ming Lin
Aug 20, 2016·Rapid Communications in Mass Spectrometry : RCM·Jing WuJin-Ming Lin
Dec 10, 2014·Analytical Chemistry·Bernhard Spengler
Mar 9, 2013·Chemical Society Reviews·Mathivathani Kandiah, Pawel L Urban
May 2, 2015·Electrophoresis·Petra Dittrich, Alfredo J Ibáñez
Dec 31, 2018·Applied Microbiology and Biotechnology·Oluwaseyi Samuel Olanrewaju, Olubukola Oluranti Babalola

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