Electroporation and mass spectrometry: a new paradigm for in situ analysis of intact proteins direct from living yeast colonies

Research output: Contribution to journalArticlepeer-review

Standard

Electroporation and mass spectrometry : a new paradigm for in situ analysis of intact proteins direct from living yeast colonies. / Kocurek, Klaudia; Havlikova, Jana; Buchan, Emma; Tanner, Andrew; May, Robin; Cooper, Helen.

In: Analytical Chemistry, Vol. 92, No. 3, 04.02.2020, p. 2605-2611.

Research output: Contribution to journalArticlepeer-review

Harvard

APA

Vancouver

Author

Bibtex

@article{166a6e04bc184ca6911de664b2739ec8,
title = "Electroporation and mass spectrometry: a new paradigm for in situ analysis of intact proteins direct from living yeast colonies",
abstract = "Yeasts constitute an oft-neglected class of pathogens among which the resistance to first-line treatments, attributed in part to mutations in efflux pumps, is rapidly emerging. Their thick, chitin-reinforced cell walls render cell lysis difficult, complicating their analysis and identification by methods routinely used for bacteria, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Liquid extraction surface analysis mass spectrometry (LESA-MS) has previously been applied to the analysis of intact proteins from Gram-positive and Gram-negative bacterial colonies sampled directly on solid nutrient media. To date, a similar analysis of yeast colonies has not proved possible. Here we demonstrate the rapid release of intact yeast proteins for LESA-MS by electroporation using a home-built high-voltage device designed to lyse cells grown in colonies on agar media. Detection and identification of previously inaccessible proteins from baker{\textquoteright}s yeast Saccharomyces cerevisiae, as well as two clinically relevant yeast species (Candida glabrata and Cryptococcus neoformans), is shown. The electroporation approach also has the potential to be translated to other mass spectrometric analysis techniques, including MALDI and various ambient ionization methods.",
author = "Klaudia Kocurek and Jana Havlikova and Emma Buchan and Andrew Tanner and Robin May and Helen Cooper",
year = "2020",
month = feb,
day = "4",
doi = "10.1021/acs.analchem.9b04365",
language = "English",
volume = "92",
pages = "2605--2611",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "3",

}

RIS

TY - JOUR

T1 - Electroporation and mass spectrometry

T2 - a new paradigm for in situ analysis of intact proteins direct from living yeast colonies

AU - Kocurek, Klaudia

AU - Havlikova, Jana

AU - Buchan, Emma

AU - Tanner, Andrew

AU - May, Robin

AU - Cooper, Helen

PY - 2020/2/4

Y1 - 2020/2/4

N2 - Yeasts constitute an oft-neglected class of pathogens among which the resistance to first-line treatments, attributed in part to mutations in efflux pumps, is rapidly emerging. Their thick, chitin-reinforced cell walls render cell lysis difficult, complicating their analysis and identification by methods routinely used for bacteria, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Liquid extraction surface analysis mass spectrometry (LESA-MS) has previously been applied to the analysis of intact proteins from Gram-positive and Gram-negative bacterial colonies sampled directly on solid nutrient media. To date, a similar analysis of yeast colonies has not proved possible. Here we demonstrate the rapid release of intact yeast proteins for LESA-MS by electroporation using a home-built high-voltage device designed to lyse cells grown in colonies on agar media. Detection and identification of previously inaccessible proteins from baker’s yeast Saccharomyces cerevisiae, as well as two clinically relevant yeast species (Candida glabrata and Cryptococcus neoformans), is shown. The electroporation approach also has the potential to be translated to other mass spectrometric analysis techniques, including MALDI and various ambient ionization methods.

AB - Yeasts constitute an oft-neglected class of pathogens among which the resistance to first-line treatments, attributed in part to mutations in efflux pumps, is rapidly emerging. Their thick, chitin-reinforced cell walls render cell lysis difficult, complicating their analysis and identification by methods routinely used for bacteria, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Liquid extraction surface analysis mass spectrometry (LESA-MS) has previously been applied to the analysis of intact proteins from Gram-positive and Gram-negative bacterial colonies sampled directly on solid nutrient media. To date, a similar analysis of yeast colonies has not proved possible. Here we demonstrate the rapid release of intact yeast proteins for LESA-MS by electroporation using a home-built high-voltage device designed to lyse cells grown in colonies on agar media. Detection and identification of previously inaccessible proteins from baker’s yeast Saccharomyces cerevisiae, as well as two clinically relevant yeast species (Candida glabrata and Cryptococcus neoformans), is shown. The electroporation approach also has the potential to be translated to other mass spectrometric analysis techniques, including MALDI and various ambient ionization methods.

U2 - 10.1021/acs.analchem.9b04365

DO - 10.1021/acs.analchem.9b04365

M3 - Article

C2 - 31922714

VL - 92

SP - 2605

EP - 2611

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 3

ER -