Snake venomics: Characterization of protein families in Sistrurus barbouri venom by cysteine mapping, N‐terminal sequencing, and tandem mass spectrometry analysis

Juárez, Paula; Sanz, Libia; Calvete, Juan J.

doi:10.1002/pmic.200300628

Cited by 125 publications

(133 citation statements)

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“…Scatterplot of individual ion events m/z versus the normalized kinetic energy of each impact (c). Numbers in brackets indicate proteins shown iníTableí1íandípreviouslyíidentifiedí [14]. spots is cumbersome and very dependent of the capability of the protein species to bind the chromogenic dye usedíforístainingí [16].íAsíexamples,ítheíproteiní4íofí48.5 kDa (PIII metalloprotease) seems to be the most abundant protein in our analysis, both in the mass spectrum andíRP-HPLCí(Figuresí1aíandí2a).íThisíproteinírepre-sents 5.3% of the total number of ions detected by cryodetector MALDI-TOF.…”

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

“…Isolated proteins could be unambiguously assigned to protein families present in the Swiss-Prot/Tr EMBLídatabaseí [14].íSomeírepresentativeíexamplesíare includedíiníTableí1.íSeparationíofítheíproteinícompo-nents of the crude venom extract by 2D gel revealed the presenceíofífouríorífiveímajoríproteiníspotsí(Figureí2b), and the presence of isoforms for the serine protease (proteiní2)í [14].…”

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

“…The crude venom of Sistrurus miliarius barbouri was fractionatedí byí reverse-phaseí HPLCí (Figureí 2a).í The characterization of the proteins present in each fractioníwasípreviouslyídoneíbyíJuarezíetíal.í [14],íusing SDS-PAGE, MALDI-TOF MS, Edman degradation (N-terminal sequencing), and tandem mass spectrometry in a quadrupole-linear ion trap instrument (data not shown). Isolated proteins could be unambiguously assigned to protein families present in the Swiss-Prot/Tr EMBLídatabaseí [14].íSomeírepresentativeíexamplesíare includedíiníTableí1.íSeparationíofítheíproteinícompo-nents of the crude venom extract by 2D gel revealed the presenceíofífouríorífiveímajoríproteiníspotsí(Figureí2b), and the presence of isoforms for the serine protease (proteiní2)í [14].…”

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

“…High-mass cryodetection (a) and microchannel plate (b) MALDI-TOF mass spectra of the S. miliarius barbouri venom extract. 0.8 g of total protein was deposited onto the MALDI plate without previousí fractionation.í Numbersí iní bracketsí indicateí proteinsí showní iní Tableí 1í andí previously identifiedí [14]. time of the different ions.…”

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

“…The results were compared with those obtainedíbyíJuarezíetíal.í [14]íusingí2Dígelíelectrophoresis and reversed-phase HPLC liquid chromatography (LC), highlighting the sensitivity of cryodetection and its ability for relative quantification of different analytes in the same spectrum. Overall, we demonstrate the power of this new technology to be implemented as a mass spectrometrybased quantitative proteomic profiling strategy.…”

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Proteomic profiling of a snake venom using high mass detection MALDI-TOF mass spectrometry

Yanes

Avilés

Wenzel

et al. 2007

J. Am. Soc. Mass Spectrom.

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Proteomic profiling involves identification and quantification of protein components in complex biological systems. Most of the mass profiling studies performed with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) have been restricted to peptides and small proteins (Ͻ20 kDa) because the sensitivity of the standard ion detectors decreases with increasing ion mass. Here we perform a protein profiling study of the snake venom Sistrurus miliarius barbouri, comparing 2D gel electrophoresis and reversed-phase high-performance liquid chromatography (HPLC) with a high mass cryodetector MALDI-TOF instrument (Macromizer), whose detector displays an uniform sensitivity with mass. Our results show that such MS approach can render superior analysis of protein complexity compared with that obtained with the electrophoretic and chromatographic approaches. The summation of ion impacts allows relative quantification of different proteins, and the number of ion counts correlates with the peak areas in the reversed-phase HPLC. Furthermore, the sensitivity reached with the high mass cryodetection MS technology clearly exceeds the detection limit of standard high-sensitivity staining methods. (J Am Soc Mass Spectrom 2007, 18, 600 -606) © 2007 American Society for Mass Spectrometry P rotein profiling has become a powerful method for analyzing changes in global protein expression patterns in biological systems as a function of developmental, physiological, and disease processes. Two-dimensional (2D) gel electrophoresis has been established for many years as the primary tool for detecting proteins present in an organism or a complex biological extract. However, 2D gel electrophoresis has some limitations: limited solubility of hydrophobic and membrane proteins, limited dynamic range, difficulty in focusing highly basic and acidic proteins, poor sensitivity, poor quantitation, and finally, the method is not amenable to automation. The limitations associated with gel electrophoretic analysis of peptides and small proteins (Ͻ20 kD) have stimulated interest in mass spectrometry as an alternative strategy. In this sense, MALDI-TOF mass spectrometry has partially replaced or complemented gel-based approaches for studying the proteomic profile of the low mass components of many complex biological samples such as serum [1] or tissues [2]. Imaging MS [2] and chip-based technologies such as surfaceenhanced laser desorption/ionization (SELDI-MS) [3] are relatively new approaches that take advantage of the established features of MALDI MS (mass accuracy, sensitivity, reliability, and high-throughput) to detect and identify peptides and proteins. Also, 1D and 2D liquid chromatography coupled to detection systems such as UV and/or MALDI, have been gradually introduced for the analysis and profiling of intact (top down proteomics) [4,5] and digested proteins (bottom up proteomics) [6] in complex biological mixtures. MALDI is a soft ionization method and produces predominantly singly charged molecular ions r...

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Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

Section: Quantitative Profiling Of the Snake Venommentioning

confidence: 99%

mentioning

confidence: 99%

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Proteomic profiling of a snake venom using high mass detection MALDI-TOF mass spectrometry

Yanes

Avilés

Wenzel

et al. 2007

J. Am. Soc. Mass Spectrom.

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Current awareness on comparative and functional genomics

2004

Comp Funct Genom

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on comparative and functional genomics. Each bibliography is divided into 16 sections. 1 Reviews & symposia; 2 General; 3 Large‐scale sequencing and mapping; 4 Genome evolution; 5 Comparative genomics; 6 Gene families and regulons; 7 Pharmacogenomics; 8 Large‐scale mutagenesis programmes; 9 Functional complementation; 10 Transcriptomics; 11 Proteomics; 12 Protein structural genomics; 13 Metabolomics; 14 Genomic approaches to development; 15 Technological advances; 16 Bioinformatics. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted

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Fractionation and proteomic analysis of the Walterinnesia aegyptia snake venom using OFFGEL and MALDI‐TOF‐MS techniques

El-Aziz

Bourgoin-Voillard

Combemale³

et al. 2015

Electrophoresis

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Animal venoms are complex mixtures of more than 100 different compounds, including peptides, proteins, and nonprotein compounds such as lipids, carbohydrates, and metal ions. In addition, the existing compounds show a wide range of molecular weights and concentrations within these venoms, making separation and purification procedures quite tedious. Here, we analyzed for the first time by MS the advantages of using the OFFGEL technique in the separation of the venom components of the Egyptian Elapidae Walterinnesia aegyptia snake compared to two classical methods of separation, SEC and RP-HPLC. We demonstrate that OFFGEL separates venom components over a larger scale of fractions, preserve respectable resolution with regard to the presence of a given compound in adjacent fractions and allows the identification of a greater number of ions by MS (102 over 134 total ions). We also conclude that applying several separating techniques (SEC and RP-HPLC in addition to OFFGEL) provides complementary results in terms of ion detection (21 more for SEC and 22 more with RP-HPLC). As a result, we provide a complete list of 134 ions present in the venom of W. aegyptia by using all these techniques combined.

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Snake venomics: Characterization of protein families in Sistrurus barbouri venom by cysteine mapping, N‐terminal sequencing, and tandem mass spectrometry analysis

Cited by 125 publications

References 39 publications

Proteomic profiling of a snake venom using high mass detection MALDI-TOF mass spectrometry

Proteomic profiling of a snake venom using high mass detection MALDI-TOF mass spectrometry

Current awareness on comparative and functional genomics

Fractionation and proteomic analysis of the Walterinnesia aegyptia snake venom using OFFGEL and MALDI‐TOF‐MS techniques

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