Electroporation and mass spectrometry: a new paradigm for in situ analysis of intact proteins direct from living yeast colonies
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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 journal › Article › peer-review
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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 -