QuantiSpec--Quantitative mass spectrometry data analysis of (15)N-metabolically labeled proteins.
J Proteomics
; 71(6): 601-8, 2009 Jan 30.
Article
em En
| MEDLINE
| ID: mdl-19028610
ABSTRACT
For relative protein quantitation by mass spectrometry we metabolically labeled E. coli bacteria with (15)N-enriched diets. Proteins extracted from (15)N-labeled and unlabeled E. coli bacteria were mixed, separated by two-dimensional gel electrophoresis and enzymatically digested. The resulting tryptic peptides were analyzed by MALDI mass spectrometry. For the relative protein quantitation we developed fully automated software, QuantiSpec (Quantitative Mass Spectrometry Analysis Software), which uses data from MALDI TOF mass spectrometry and the Mascot database search engine. QuantiSpec detects natural as well as partially or fully labeled peptide isotope distributions. For each identified peptide the (15)N incorporation rate is determined by comparing the experimental to a set of theoretical isotope patterns based on the peptide sequence. Relative quantitation is accomplished by calculating the signal intensity ratios for each (14)N/(15)N peptide pair.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteoma
/
Proteínas de Escherichia coli
/
Isótopos de Nitrogênio
Idioma:
En
Revista:
J Proteomics
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
Alemanha