About: Evaluation of the possible proteomic application of trypsin from Streptomyces griseus

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	In this work, trypsin from Streptomyces griseus (SGT) was purified to homogeneity and evaluated in in-gel digestion of protein standards followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analyses of the digests. The number of produced and matching tryptic peptides was higher than in the case of commonly used bovine trypsin and allowed us to obtain higher identification scores in database searches. Interestingly, SGT was found to also generate nonspecific peptides, a partial F-X, Y-X, and W-X cleavage specificity. To suppress autolysis, either arginine or arginine plus lysine residues in SGT were modified by chemical reagents. In consequence, the autolytic pattern of SGT was reduced significantly, but specific activity dropped dramatically. As demonstrated by relative quantification, SGT is more stable at 37 degs than is its bovine counterpart. We conclude that SGT represents a convenient alternative for proteomic applications involving protein digestion. In this work, trypsin from Streptomyces griseus (SGT) was purified to homogeneity and evaluated in in-gel digestion of protein standards followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analyses of the digests. The number of produced and matching tryptic peptides was higher than in the case of commonly used bovine trypsin and allowed us to obtain higher identification scores in database searches. Interestingly, SGT was found to also generate nonspecific peptides, a partial F-X, Y-X, and W-X cleavage specificity. To suppress autolysis, either arginine or arginine plus lysine residues in SGT were modified by chemical reagents. In consequence, the autolytic pattern of SGT was reduced significantly, but specific activity dropped dramatically. As demonstrated by relative quantification, SGT is more stable at 37 degs than is its bovine counterpart. We conclude that SGT represents a convenient alternative for proteomic applications involving protein digestion. (en)
Title	Evaluation of the possible proteomic application of trypsin from Streptomyces griseus Evaluation of the possible proteomic application of trypsin from Streptomyces griseus (en)
skos:prefLabel	Evaluation of the possible proteomic application of trypsin from Streptomyces griseus Evaluation of the possible proteomic application of trypsin from Streptomyces griseus (en)
skos:notation	RIV/00216224:14310/08:00025879!RIV10-MSM-14310___
http://linked.open...avai/riv/aktivita	P S Z
http://linked.open...avai/riv/aktivity	P(LC06034), S, Z(AV0Z40310501), Z(MSM0021622413), Z(MSM0021622415), Z(MSM6198959216)
http://linked.open...iv/cisloPeriodika	1
http://linked.open...vai/riv/dodaniDat	2010
http://linked.open...aciTvurceVysledku	Šebela, Marek Řehulka, Pavel Zdráhal, Zbyněk Šedo, Ondrej Havliš, Jan
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Masarykova univerzita / Přírodovědecká fakulta
http://linked.open...dnocenehoVysledku	366864
http://linked.open...ai/riv/idVysledku	RIV/00216224:14310/08:00025879
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	In gel digestion; MALDI TOF MS; Peptide mass fingerprinting; Protein modification; Streptomyces griseus; Trypsin (en)
http://linked.open.../riv/klicoveSlovo	MALDI TOF MS Trypsin In gel digestion Peptide mass fingerprinting Protein modification Streptomyces griseus
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[C111382F026A]
http://linked.open...i/riv/nazevZdroje	Analytical Biochemistry
http://linked.open...in/vavai/riv/obor	CE
http://linked.open...ichTvurcuVysledku	5 (xsd:int)
http://linked.open...cetTvurcuVysledku	6 (xsd:int)
http://linked.open...vavai/riv/projekt	Regulation of morphogenesis of plant cells and organs
http://linked.open...UplatneniVysledku	2008
http://linked.open...v/svazekPeriodika	376
http://linked.open...iv/tvurceVysledku	Zdráhal, Zbyněk Šedo, Ondrej Šebela, Marek Řehulka, Pavel Havliš, Jan Štosová, Tatiana
http://linked.open...ain/vavai/riv/wos	000255104100009
http://linked.open...n/vavai/riv/zamer	Moderní analytické techniky pro bioanalýzu, ekologii a nanotechnologie Proteiny v metabolismu a při interakci organismů s prostředím Molecular basis of cell and tissue regulations Modulation of signalling and regulatory pathways in normal and cancer cells
issn	0003-2697
number of pages	9 (xsd:int)
http://localhost/t...ganizacniJednotka	14310

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