2,5-Dihydroxybenzoic acid instead of α-cyano-4-hydroxycinnamic acid as matrix in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for analyses of in-gel digests of silver-stained proteins

Ghafouri, Bijar; Karlsson, Helén; Mörtstedt, Harriet; Lewander, Andreas; Tagesson, Christer; Lindahl, Mattias

doi:10.1016/j.ab.2007.07.002

Cited by 15 publications

(18 citation statements)

References 13 publications

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“…Protein spots of interest were excised from the gel and, after tryptic digestion, were analyzed by mass spectrometry using ultrafleXtreme™ matrix-assisted laser desorption/ionization – time of flight (MALDI-TOF; Bruker Daltronik GmbH, Bremen, Germany). Database search was performed in ProteinProspector MS-Fit version 5.14.4 including Swiss-Prot database version 2015.3.5 as described in previous studies 28,32…”

Section: Methodsmentioning

confidence: 99%

Systemic alterations in plasma proteins from women with chronic widespread pain compared to healthy controls: a proteomic study

Wåhlén¹,

Olausson²,

Carlsson³

et al. 2017

JPR

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Chronic widespread pain (CWP) is a complex pain condition that is difficult to treat. The prevalence of CWP approximates ~10% of the general population, with higher prevalence in women. Lack of understanding of molecular mechanisms has been a challenge for diagnosis and treatment of chronic pain. The aim of this study was to explore the systemic protein changes in CWP compared to those in healthy controls (CON). By applying 2-dimensional gel electrophoresis, we analyzed the protein pattern of plasma samples from women with CWP (n=16) and healthy women (n=23). The proteomic data were analyzed using multivariate statistical models, and altered proteins were identified using mass spectrometry. The proteome analysis was further validated by gel-free Western blot. Multivariate statistical data analysis of quantified proteins revealed 22 altered proteins in women with CWP, compared to CON group. Many of the identified proteins are previously known to be involved in different parts of the complement system and metabolic and inflammatory processes, e.g., complement factor B, vitamin D-binding protein, ceruloplasmin, transthyretin and alpha-2-HS-glycoprotein. These results indicate that important systemic protein differences exist between women with CWP and healthy women. Further, this study illustrates the potential use of proteomics to detect biomarkers that may provide new insights into the molecular mechanism(s) of chronic pain. However, further larger investigations are required in order to confirm these findings before it will be possible to identify proteins as potential pain biomarkers for clinical use.

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Section: Methodsmentioning

confidence: 99%

Systemic alterations in plasma proteins from women with chronic widespread pain compared to healthy controls: a proteomic study

Wåhlén¹,

Olausson²,

Carlsson³

et al. 2017

JPR

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“…Trypsin (10 mg/ml Sequencing Grade Modified, Promega, WI, USA) was added to the samples, incubated over night in 37° C and then dried in speed-vac. The peptides were reconstituted in 0.1 % trifluoroacetic acid and mixed 1:1 with matrix (2,5-dihydroxybenzoic acid, 0.02 mg/ml) as described previously (Ghafouri et al 2007). The sample was applied onto the target plate (V700666 REV.C, Applied Biosystems) and the peptide masses were acquired using a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS, Voyager-DE PRO, Applied Biosystems, Foster City, CA, USA) using a 337 nm N 2 laser, delayed extraction, positive ionization, operated in reflector mode and instrument settings as defined earlier (Ghafouri et al 2007).…”

Section: Identification Of Proteins With Mass Spectrometrymentioning

confidence: 99%

“…The peptides were reconstituted in 0.1 % trifluoroacetic acid and mixed 1:1 with matrix (2,5-dihydroxybenzoic acid, 0.02 mg/ml) as described previously (Ghafouri et al 2007). The sample was applied onto the target plate (V700666 REV.C, Applied Biosystems) and the peptide masses were acquired using a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS, Voyager-DE PRO, Applied Biosystems, Foster City, CA, USA) using a 337 nm N 2 laser, delayed extraction, positive ionization, operated in reflector mode and instrument settings as defined earlier (Ghafouri et al 2007). Spectra in the mass range from 700 to 3600 m/z were collected from 400 laser pulses/sample with close external mass calibration with a standard peptide mixture and internal calibration using known trypsin autolysis peaks (m/z 842.5100, 2211.1046).…”

Section: Identification Of Proteins With Mass Spectrometrymentioning

confidence: 99%

Airway irritation among indoor swimming pool personnel: trichloramine exposure, exhaled NO and protein profiling of nasal lavage fluids

Fornander

Ghafouri

Lindahl

et al. 2012

Int Arch Occup Environ Health

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Purpose Occurrence of airway irritation among indoor swimming pool personnel was investigated.The aim was to assess trichloramine exposure levels and exhaled nitric oxide in relation to the prevalence of airway symptoms in swimming pool facilities and to determine protein effects in the upper respiratory tract. MethodsThe presence of airway symptoms related to work were examined in 146 individuals working at 46 indoor swimming pool facilities. Levels of trichloramine, as well as exhaled nitric oxide, were measured in five facilities with high prevalence of airway irritation and four facilities with no airway irritation among the personnel. Nasal lavage fluid was collected and protein profiles were determined by a proteomic approach.Results 17 % of the swimming pool personnel reported airway symptoms related to work. The levels of trichloramine in the swimming pool facilities ranged from 0.04-0.36 mg/m 3 . There were no covariance between trichloramine levels, exhaled nitric oxide and prevalence of airway symptoms.Protein profiling of the nasal lavage fluid showed that, alpha-1-antitrypsin and lactoferrin, were significantly higher, and S100-A8 was significantly lower in swimming pool personnel.Conclusions This study confirms the occurrence of airway irritation among indoor swimming pool personnel. Our results indicate altered levels of innate immunity proteins in the upper airways that may pose as potential biomarkers. However, swimming pool facilities with high prevalence of airway irritation could not be explained by higher trichloramine exposure levels. Further studies are needed to clarify the environmental factors in indoor swimming pools that cause airway problems and affect the immune system.

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“…The far most commonly used matrix for peptide mass fingerprinting is CHCA, which is recommended for peptides with mass ions below 2500 Da (Beavis et al, 1992). Alternative matrices also used are sinapinic acid, mostly for masses higher than 25 kDa (Lewis et al, 2000) and DHB, originally suggested for glyco-or phosphopepides that are difficult to ionize (Strupat et al, 1991), but later also proven useful for silver stained proteins (Ghafouri et al, 2007). As illustrated in figure 11, CHCA and DHB have very different crystal structures on the target plate.…”

Section: Choice Of Matrixmentioning

confidence: 99%

Two-Dimensional Gel Electrophoresis and Mass Spectrometry in Studies of Nanoparticle-Protein Interactions

Karlsson¹,

Ljunggren²,

Ahrén³

et al. 2012

Gel Electrophoresis - Advanced Techniques

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2,5-Dihydroxybenzoic acid instead of α-cyano-4-hydroxycinnamic acid as matrix in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for analyses of in-gel digests of silver-stained proteins

Cited by 15 publications

References 13 publications

Systemic alterations in plasma proteins from women with chronic widespread pain compared to healthy controls: a proteomic study

Systemic alterations in plasma proteins from women with chronic widespread pain compared to healthy controls: a proteomic study

Airway irritation among indoor swimming pool personnel: trichloramine exposure, exhaled NO and protein profiling of nasal lavage fluids

Two-Dimensional Gel Electrophoresis and Mass Spectrometry in Studies of Nanoparticle-Protein Interactions

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