Desorption electrospray ionization mass spectrometry of intact bacteria

Meetani, Mohammed A.; Shin, Yun‐Jeong; Zhang, Shaofeng; Mayer, Richard T.; Basile, Franco

doi:10.1002/jms.1250

Cited by 78 publications

(63 citation statements)

References 44 publications

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“…Third, the last decade has seen very significant advances in MS with respect to ion sources and the sensitivity of the instruments themselves (20)(21)(22)(23)(24)(25)(26)(27). Ambient ionization methods combined with significant improvements in sensitivity and mass accuracy of MS instrumentation now enable the detection of intact molecules directly from surfaces (7,(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40). Using the ambient method nanoDESI, the molecular characterization of microbial colonies directly from agar surfaces without any prior sample preparation has become possible (7).…”

Section: Significancementioning

confidence: 99%

MS/MS networking guided analysis of molecule and gene cluster families

Nguyen

Moree

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

235

238

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The ability to correlate the production of specialized metabolites to the genetic capacity of the organism that produces such molecules has become an invaluable tool in aiding the discovery of biotechnologically applicable molecules. Here, we accomplish this task by matching molecular families with gene cluster families, making these correlations to 60 microbes at one time instead of connecting one molecule to one organism at a time, such as how it is traditionally done. We can correlate these families through the use of nanospray desorption electrospray ionization MS/MS, an ambient pressure MS technique, in conjunction with MS/MS networking and peptidogenomics. We matched the molecular families of peptide natural products produced by 42 bacilli and 18 pseudomonads through the generation of amino acid sequence tags from MS/MS data of specific clusters found in the MS/MS network. These sequence tags were then linked to biosynthetic gene clusters in publicly accessible genomes, providing us with the ability to link particular molecules with the genes that produced them. As an example of its use, this approach was applied to two unsequenced Pseudoalteromonas species, leading to the discovery of the gene cluster for a molecular family, the bromoalterochromides, in the previously sequenced strain P. piscicida JCM 20779 T . The approach itself is not limited to 60 related strains, because spectral networking can be readily adopted to look at molecular family-gene cluster families of hundreds or more diverse organisms in one single MS/MS network.MS/MS molecular networking | mass spectrometry | microbial ecology T ens of thousands of sequenced microbial genomes or rough drafts of genomes are available at this time, and this number is predicted to grow into the millions over the next decades. This wealth of sequence data has the potential to be used for the discovery of small bioactive molecules through genome mining (1-6). Genome mining is a process in which small molecules are discovered by predicting what compound will be genetically encoded based on the sequences of biosynthetic gene clusters. However, the process of mining genetically encoded small molecules is not keeping pace with the rate by which genome sequences are being obtained. In general, genome mining is still done one gene cluster at a time and requires many person-years of effort to annotate a single molecule. The time and significant expertise that current genome mining requires also make genome mining very expensive. In light of this extensive effort and cost, alternative approaches to genome mining and annotating specialized metabolites must be developed that not only take advantage of the sequenced resources available and make it efficient to perform genome mining on a more global scale but also enable the molecular analysis of unsequenced organisms. Such methods will then significantly reduce the cost of genome mining by increasing the speed with which molecules are connected to candidate genes and using resources already available. Here, we put fo...

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Section: Significancementioning

confidence: 99%

MS/MS networking guided analysis of molecule and gene cluster families

Nguyen

Moree

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

235

238

View full text Add to dashboard Cite

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“…For example, MS analysis and identification of microorganisms, mostly performed by commercial platforms based on MALDI, is revolutionizing the field of microbiology [30]. Similarly, using DESI, direct analysis of microorganisms showed that characteristic constituents of bacteria can be detected; in particular, acylium ions of FA are observable directly, not as the usual methyl ester derivatives [32,33]. DESI allows rapid in situ analysis of samples in their native state and has been used to study the chemical composition of animal tissues, plants, and documents in an untargeted fashion [34].…”

Section: Introductionmentioning

confidence: 99%

Developmental phases of individual mouse preimplantation embryos characterized by lipid signatures using desorption electrospray ionization mass spectrometry

et al. 2012

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Knowledge of the lipids present in individual preimplantation embryos is of interest in fundamental studies of embryology, in attempts to understand cellular pluripotency and in optimization of in vitro culture conditions necessary for the application and development of biotechnologies such as in vitro fertilization and transgenesis. In this work, the profiles of fatty acids and phospholipids (PL) in individual mouse preimplantation embryos and oocytes were acquired using an analytical strategy based on desorption electrospray ionization mass spectrometry (DESI-MS). The methodology avoids sample preparation and provides information on the lipids present in these microscopic structures. Differences in the lipid profiles observed for unfertilized oocytes, two-and four-cell embryos, and blastocysts were characterized. For a representative set of embryos (N= 114) using multivariate analysis (specifically principal component analysis) unfertilized oocytes showed a narrower range of PL species than did blastocysts. Two-and four-cell embryos showed a wide range of PLs compared with unfertilized oocytes and high abundances of fatty acids, indicating pronounced synthetic activity. The data suggest that the lipid changes observed in mouse preimplantation development reflect acquisition of a degree of cellular membrane functional and structural specialization by the blastocyst stage. It is also noteworthy that embryos cultured in vitro from the two-cell through the blastocyst stage have a more homogeneous lipid profile as compared with their in vivo-derived counterparts, which is ascribed to the restricted diversity of nutrients present in synthetic culture media. The DESI-MS data are interpreted from lipid biochemistry and previous reports on gene expression of diverse lipids known to be vital to early embryonic development.

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“…Although GC-MS instruments currently dominate the portable mass spectrometry industry, estimates indicate that the confidence for molecule identification by MS/MS is about that provided with GC-MS (when combining MS information with retention time) [15]. The MS/MS-capable CITbased instrument enables chromatography-free analysis for high confidence in situ molecule identification with 100-fold faster analysis times than standard GC-MS analyses.Atmospheric pressure desorption ionization techniques, including DESI and DART, are capable of generating ions of diverse chemical nature directly from solid surfaces for MS/MS analysis [10,[16][17][18][19][20][21][22][23][24][25][26][27][28][29][30]. In forensic [26,31], security [19,32], and quality control applications [17,27], desorption techniques provide actionable chemical information through rapid, definitive analyses.…”

mentioning

confidence: 99%

Implementation of DART and DESI ionization on a fieldable mass spectrometer

Wells¹,

Roth²,

Keil³

et al. 2008

J. Am. Soc. Mass Spectrom.

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A recently developed prototype mobile laboratory mass spectrometer, incorporating an atmospheric pressure ionization (API) interface, is described. This system takes advantage of the small size, lower voltage requirements, and tandem MS abilities of the cylindrical ion trap mass analyzer. The prototype API MS uses small, low-power pumps to fit into a 0.1-m 3 self-contained package weighing <45 kg. This instrument has been adapted to allow rapid interfacing to electrospray ionization, desorption electrospray ionization, and direct analysis in real-time sources. Initial data indicate that these techniques provide rapid detection and identification of compounds for quality control, homeland security, and forensic applications. In addition, this instrument is self-contained and compact, making it ideally extensible to mobile laboratory and field analyses. Initial MS and MS/MS data for analyses of drugs, food, and explosives are presented herein. O ver the past 15 years, efforts toward implementing mass spectrometry (MS) in the field have steadily grown with particular focus on environmental, forensic, defense, and security applications [I, 2]. Many of the previously developed fieldable MS instruments use gas chromatography (GC-MS) because of the high degree of confidence obtained using GC retention times and electron ionization (EI) mass spectral matching [3][4][5][6]. The specificity of GC-MS comes at the expense of time, with individual analyses often taking in excess of 10 min (not including sample preparation). Reducing the amount of time required for chemical identification requires separation-free MS. In the absence of chromatography, tandem MS (MS/MS) improves confidence for the identification of individual components in mixtures [7]. With spectral acquisition rates > 1 Hz, atmospheric pressure ionization (API) techniques such as electrospray ionization (ESI), desorption electro spray (DESI), and direct analysis in real time (DART), a fieldable API MS system capable of MS/MS is of high value to environmental, forensic, defense, and force protection agencies. There have been few attempts to construct fieldable MS instruments with an API interface to date [8][9][10][11] because of the difficulty of displacing the gas load of the atmospheric inlet.The cylindrical ion trap (CIT)-a simplified geometry of the hyperbolic quadrupole ion trap capable of Address reprint requests to Dr. Mitch Wells, Griffin Analytical Technologies, LLC, R&D, 3000 Kent Avenue, West Lafayette, IN 47906, USA. E-mail: mitch.wells@icxt.com performing MS n analyses-has been shown to be amenable to miniaturization [12][13][14] while maintaining benchtop-quality performance characteristics. The miniaturized CIT enables a smaller vacuum system with smaller pumps that consume less power, providing a fieldable instrument. Although GC-MS instruments currently dominate the portable mass spectrometry industry, estimates indicate that the confidence for molecule identification by MS/MS is about that provided with GC-MS (when combining MS information w...

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Desorption electrospray ionization mass spectrometry of intact bacteria

Cited by 78 publications

References 44 publications

MS/MS networking guided analysis of molecule and gene cluster families

MS/MS networking guided analysis of molecule and gene cluster families

Developmental phases of individual mouse preimplantation embryos characterized by lipid signatures using desorption electrospray ionization mass spectrometry

Implementation of DART and DESI ionization on a fieldable mass spectrometer

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