Detailed Information on Publication Record
2008
Evaluation of the possible proteomic application of trypsin from Streptomyces griseus
HAVLIŠ, Jan, Tatiana ŠTOSOVÁ, Marek ŠEBELA, Pavel ŘEHULKA, Ondrej ŠEDO et. al.Basic information
Original name
Evaluation of the possible proteomic application of trypsin from Streptomyces griseus
Name in Czech
Zjištění možných proteomických uplatnění tryspinu ze Streptomyces griseus
Authors
HAVLIŠ, Jan (203 Czech Republic, belonging to the institution), Tatiana ŠTOSOVÁ (203 Czech Republic), Marek ŠEBELA (203 Czech Republic, guarantor), Pavel ŘEHULKA (203 Czech Republic), Ondrej ŠEDO (203 Czech Republic, belonging to the institution) and Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution)
Edition
Analytical Biochemistry, San Diego, Academic Press, 2008, 0003-2697
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10600 1.6 Biological sciences
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
Impact factor
Impact factor: 3.088
RIV identification code
RIV/00216224:14310/08:00025879
Organization unit
Faculty of Science
UT WoS
000255104100009
Keywords in English
In gel digestion; MALDI TOF MS; Peptide mass fingerprinting; Protein modification; Streptomyces griseus; Trypsin
Tags
Tags
International impact, Reviewed
Změněno: 29/3/2013 09:55, Mgr. Ondrej Šedo, Ph.D.
V originále
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. Moreover, parallel digestions of sample aliquots by SGT and BT provide the possibility of combining partially different results (unique matching peptides) to improve protein identification.
In Czech
V této práci byla popsána izolace a charakteriza trypsinu ze Streptomyces griseus pro použití v proteomice.
Links
| LC06034, research and development project |
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| MSM0021622413, plan (intention) |
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| MSM0021622415, plan (intention) |
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