scholarly journals Direct Imaging and Identification of Proteoforms up to 70 kDa from Human Tissue

2021 ◽  
Author(s):  
Pei Su ◽  
John P. McGee ◽  
Kenneth R. Durbin ◽  
Michael A. R. Hollas ◽  
Manxi Yang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Imaging Mass Spectrometry ◽  
Desorption Electrospray Ionization ◽  
Human Kidney ◽  
Primary Metabolism ◽  
Label Free ◽  
Proteome Coverage ◽  
Individual Ion ◽  
Detection And Identification ◽  
Ion Mass Spectrometry

AbstractImaging of proteoforms in human tissues is hindered by low molecular specificity and limited proteome coverage. Here, we introduce proteoform imaging mass spectrometry (PiMS), which increases the size limit for proteoform detection and identification by 4-fold compared to reported methods, and reveals tissue localization of proteoforms at <80 μm spatial resolution. PiMS advances proteoform imaging by combining liquid sampling (nanospray desorption electrospray ionization, nano-DESI) with ion detection using individual ion mass spectrometry (I2MS). We demonstrate the first proteoform imaging of human kidney, identifying 169 of 400 proteoforms <70 kDa using top-down mass spectrometry and database lookup from the human proteoform atlas, including dozens of key enzymes in primary metabolism. Moreover, PiMS images visualize kidney anatomical structures and cellular neighborhoods in the vasculature versus the medulla or the cortex. The benefits of PiMS are poised to increase proteome coverage for label-free protein imaging of intact tissues.TeaserNano-DESI combined with individual ion mass spectrometry generates images of proteoforms up to 70 kDa.

Spatial Metabolomics of the Human Kidney Using MALDI Trapped Ion Mobility Imaging Mass Spectrometry

2020 ◽  
Author(s):  
Elizabeth Neumann ◽  
Lukasz Migas ◽  
Jamie L. Allen ◽  
Richard Caprioli ◽  
Raf Van de Plas ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Imaging Mass Spectrometry ◽  
Structural Complexity ◽  
Human Kidney ◽  
Resolving Power ◽  
Mobility Separation ◽  
Trapped Ion ◽  
Mobility Data ◽  
Ion Mobility Separation

<div> <div> <p>Small metabolites are essential for normal and diseased biological function but are difficult to study because of their inherent structural complexity. MALDI imaging mass spectrometry (IMS) of small metabolites is particularly challenging as MALDI matrix clusters are often isobaric with metabolite ions, requiring high resolving power instrumentation or derivatization to circumvent this issue. An alternative to this is to perform ion mobility separation before ion detection, enabling the visualization of metabolites without the interference of matrix ions. Here, we use MALDI timsTOF IMS to image small metabolites at high spatial resolution within the human kidney. Through this, we have found metabolites, such as arginic acid, acetylcarnitine, and choline that localize to the cortex, medulla, and renal pelvis, respectively. We have also demonstrated that trapped ion mobility spectrometry (TIMS) can resolve matrix peaks from metabolite signal and separate both isobaric and isomeric metabolites with different localizations within the kidney. The added ion mobility data dimension dramatically increased the peak capacity for molecular imaging experiments. Future work will involve further exploring the small metabolite profiles of human kidneys as a function of age, gender, and ethnicity.</p></div></div>

Visualizing mass transport in desorption electrospray ionization using time-of-flight secondary ion mass spectrometry: a look at the geometric configuration of the spray

The Analyst ◽  
2014 ◽  
Vol 139 (22) ◽  
pp. 5868-5878 ◽  
Author(s):  
Shin Muramoto
Keyword(s):  
Mass Spectrometry ◽  
Mass Transport ◽  
Electrospray Ionization ◽  
Secondary Ion Mass Spectrometry ◽  
Time Of Flight ◽  
Desorption Electrospray Ionization ◽  
Geometric Configuration ◽  
Ion Mass Spectrometry ◽  
Secondary Ion

The desorption profile of analyte molecules desorbed by desorption electrospray ionization was imaged and characterized using time-of-flight secondary ion mass spectrometry.

scholarly journals Discrimination of Human Astrocytoma Subtypes by Lipid Analysis Using Desorption Electrospray Ionization Imaging Mass Spectrometry

2010 ◽  
Vol 122 (34) ◽  
pp. 6089-6092 ◽  
Author(s):  
Livia S. Eberlin ◽  
Allison L. Dill ◽  
Alexandra J. Golby ◽  
Keith L. Ligon ◽  
Justin M. Wiseman ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Ionization ◽  
Lipid Analysis ◽  
Imaging Mass Spectrometry ◽  
Desorption Electrospray Ionization ◽  
Human Astrocytoma

Desorption Electrospray Ionization Imaging Mass Spectrometry as a Tool for Investigating Model Prebiotic Reactions on Mineral Surfaces.

2013 ◽  
Vol 85 (3) ◽  
pp. 1276-1279 ◽  
Author(s):  
Rachel V. Bennett ◽  
H. James Cleaves ◽  
Jeffrey M. Davis ◽  
Denis A. Sokolov ◽  
Thomas M. Orlando ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Ionization ◽  
Imaging Mass Spectrometry ◽  
Desorption Electrospray Ionization ◽  
Mineral Surfaces

scholarly journals Multiplexed Mass Spectrometry of Individual Ions Improves Measurement of Proteoforms and Their Complexes

2021 ◽  
Author(s):  
Jared O. Kafader ◽  
Rafael D. Melani ◽  
Kenneth R. Durbin ◽  
Bon Ikwuagwu ◽  
Bryan P. Early ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Sample Preparation ◽  
Spectrometry Method ◽  
Mass Spectrometry Method ◽  
Individual Ion ◽  
Ion Mass Spectrometry ◽  
The Individual ◽  
Orbitrap Analyzer

Abstract Protocol for sample preparation, instrumental settings, and processing for the individual ion mass spectrometry method (I2MS) utilizing an Orbitrap analyzer.

scholarly journals DirectMS1: MS/MS-free identification of 1000 proteins of cellular proteomes in 5 minutes

2019 ◽  
Author(s):  
Mark V. Ivanov ◽  
Julia A. Bubis ◽  
Vladimir Gorshkov ◽  
Irina A. Tarasova ◽  
Lev I. Levitsky ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biomarker Discovery ◽  
Tandem Mass ◽  
Label Free ◽  
Hela Cell Line ◽  
Sequence Coverage ◽  
Proteome Coverage ◽  
Successful Attempt ◽  
Higher Sensitivity ◽  
Lengthy Analysis

AbstractProteome characterization relies heavily on tandem mass spectrometry (MS/MS) and is thus associated with instrumentation complexity, lengthy analysis time, and limited duty-cycle. It was always tempting to implement approaches which do not require MS/MS, yet, they were constantly failing in achieving meaningful depth of quantitative proteome coverage within short experimental times, which is particular important for clinical or biomarker discovery applications. Here, we report on the first successful attempt to develop a truly MS/MS-free and label-free method for bottom-up proteomics. We demonstrate identification of 1000 protein groups for a standard HeLa cell line digest using 5-minute LC gradients. The amount of loaded sample was varied in a range from 1 ng to 500 ng, and the method demonstrated 10-fold higher sensitivity compared with the standard MS/MS-based approach. Due to significantly higher sequence coverage obtained by the developed method, it outperforms all popular MS/MS-based label-free quantitation approaches.

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