Omics informatics Biotechnology Genomics Proteomics Metabolomics Genome informatics

Omics 와 informatics Biotechnology Genomics Proteomics Metabolomics Genome informatics Proteome informatics Metabolism informatics Bioinformatics Information Technology

Omics 의 관계 metabolomics Molecular machinery of cellular processes proteomics Characterization of proteome genomics Genome sequencing 1989 Peptide mass fingerprinting 1986 peptide sequencing by MS/MS

프로테옴 데이터 electrophoresis Mass spectrometry Database search 단백질 분리 펩타이드 분자량 측정 단백질 동정

단백질의 질량 스펙트럼 효소 질량분석기 MS spectrum 단백질 Protein database Protein id A B Peptide sequences A B

Mass spectrum (peptide mass fingerprint) intensity 422. 25 692. 35 1096. 59 1451. 75

Mass spectrum vs. database Mass spectrum database Protein A Protein B Protein C

Peptide mass match …. FNSTPKYIKSEGYGPREKYQSRPKFNSTPKDYN… Mass spectrum Spectrum of a protein in DB intensity FNSTPK YIK YQSRPKFNSTPKYIK 422. 25 692. 35 1096. 59 Tolerance ? 1451. 75

단백질 데이터베이스 non-redundant protein DB 효소: trypsin 0 Protein mass (0 -150 k. Da) Peptide mass (0 -12. 5 k. Da)

PMF search programs MS-Fit http: //prospector. ucsf. edu/ MASCOT http: //www. matrixscience. com/ Pept. Ident http: //www. expasy. org/ Peptide. Search http: //www. mann. embl-heidelberg. de/

MS-Fit

아미노산 분자량 아미노산 기호 MW (Da) Alanine A 71. 03711 Methionine M 131. 04049 Cysteine C 103. 00919 Asparagine N 114. 04293 Aspartic acid D 115. 02694 Proline P 97. 05276 Glutamic acid E 129. 04259 Glutamine Q 128. 05858 Phenylalanine F 147. 06841 Arginine R 156. 10111 Glycine G 57. 02146 Serine S 87. 03203 Histidine H 137. 05891 Threonine T 101. 04768 Isoleucine I 113. 08406 Valine V 99. 06841 Lysine K 128. 09496 Tryptophan W 186. 07931 Leucine L 113. 08406 tyrosine Y 163. 06333

PMF search results 통계

탄뎀 질량 스펙트럼 (MS/MS spectrum) energy 효소 1 st MS 단백질 Protein id 2 nd MS MS spectrum Peptide sequence Protein database Tandem MS spectrum

탄뎀 질량 스펙트럼 Peptide fragmentation x 3 H 2 N R 1 O C C y 3 H a 1 b 1 z 3 x 2 R 2 O N C C H H c 1 a 2 y 2 b 2 z 2 x 1 R 3 O N C C H H c 2 a 3 y 1 z 1 R 4 b 3 N C H H c 3 COOH

단백질 동정 (protein identification) intensity Peptide 의 탄뎀 질량 스펙트럼 compare intensity m/z a spectrum of a peptide in database m/z

modification K K E E T T M M P P I I M A oxidation

MS/MS Search programs MASCOT http: //www. matrixscience. com/ MS-TAG http: //prospector. ucsf. edu/ SEQUEST http: //fields. scripps. edu/sequest/

MASCOT

MASCOT output

Sequencing intensity …EDCBA …DECBA A m/z B C b 1 y 2 DE y 3 y 5 b 7 y 7 b 12 y 9 y 10 y 12 b 17 y 18 a 22

BLAST

Scores Homology search (BLAST) alignment score e-value Peptide Mass Fingerprint Number of matched peptides Coverage MOWSE score Probability based on MOWSE score Tandem mass search Cross-correlation coverage

Proteomics Trends 1. High-throughput Proteomics 2. High-throughput automated identification of proteins 3. 2. Quantitation 4. Quantitation of relative protein abundance

Proteome Database System for High-Throughput proteomics Proteome DB server Data management system Mass spectrometer instrument Mass spectrometer Mass spectrometer Mass search program database swissprot nr Web server EST user