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Chromosome 16. Marta Mendes Ignacio Casal Centro de Investigaciones Biológicas. August 2012. 250. CCD18. 1 50. Proteins to search for. 100. 75. 50. 37. 25. 20. 30 m g/ lane. 15. 10. Cell Lines CCD18 MCF7 Ramos. 1D SDS PAGE. MCF7. RAMOS. 250. 1 50. 100. 75. 250. - PowerPoint PPT Presentation
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CHROMOSOME 16Marta MendesIgnacio Casal
Centro de Investigaciones Biológicas
August 2012
ensembl gene ID Accession Gene Name Protein NameENSG00000011638 Q96B96 TMEM159 transmembrane protein 159 [Source:HGNC Symbol;Acc:30136]
ENSG00000064270 O75185 ATP2C2 ATPase, Ca++ transporting, type 2C, member 2 [Source:HGNC Symbol;Acc:29103]
ENSG00000118557 Q8TBY8 PMFBP1 polyamine modulated factor 1 binding protein 1 [Source:HGNC Symbol;Acc:17728]
ENSG00000134398 Q76MJ5 ERN2 endoplasmic reticulum to nucleus signaling 2 [Source:HGNC Symbol;Acc:16942]
ENSG00000169340 Q8N807 PDILT protein disulfide isomerase-like, testis expressed [Source:HGNC Symbol;Acc:27338]
ENSG00000204991 Q8WWL2 SPIRE2 spire homolog 2 (Drosophila) [Source:HGNC Symbol;Acc:30623]
PROTEINS TO SEARCH FOR
Cell LinesCCD18MCF7Ramos
1D SDS PAGE
250150100
755037
2520
15
10
30 mg/lane
CCD18
250150100
755037
2520
15
10
30 mg/lane
RAMOS
250150100
755037
2520
15
10
MCF7
30 mg/lane
Digestion with trypsin
High throughput
analysis
LTQ Orbitrap Velos coupled to a Proxeon nanoLC systemEQUIPMENT :
LC CONDITIONSPre column Proxeon ASY-column, L 2cm, ID 100 mm, 5 mm,
C18
Column Nano separations, NS-AC-11-dp3, L 15 cm, ID 75 mm, 3 mm, 120 A, Biosphere C18
Mobile phase A 0.1%FA/2%ACN
Mobile Phase B 0.1%FA in ACN
Flow rate 250 nl/min
Gradient Profile 160 min from 0 to 35% B
MS SYSTEMLow Resolution High Resolution
AGC MS1 3x104 1x107
AGC MS2 1x104 5x104
mscan 1 1
Maximum injection time 10 500
Isolation width 2 2
Acquisition method DDA - Top15
Resolution - 30000
Dynamic exclusion 90 s
Q8TBY8-2
CCD18MCF7
RAMOS
HIGH THROUGHPUT DDA TOP15
No proteins found
Proteome discoverer and Sequest with FDR 1%Database: uniprot_sprot_human_HPP
Peptide XCorr q-value PEP
KLEEENENLR 2.25 0.0098 0.0984
KLEEENENLR
ensembl gene ID Accession Gene Name Protein Name MW (Da) FractionsENSG00000011638 Q96B96 TMEM159 transmembrane protein 159 [Source:HGNC Symbol;Acc:30136] 17,522 10;11;12;13ENSG00000064270 O75185 ATP2C2 ATPase, Ca++ transporting, type 2C, member 2 [Source:HGNC Symbol;Acc:29103] 103,187 2;3ENSG00000118557 Q8TBY8 PMFBP1 polyamine modulated factor 1 binding protein 1 [Source:HGNC Symbol;Acc:17728] 119,033 2;3ENSG00000134398 Q76MJ5 ERN2 endoplasmic reticulum to nucleus signaling 2 [Source:HGNC Symbol;Acc:16942] 102,480 2;3ENSG00000169340 Q8N807 PDILT protein disulfide isomerase-like, testis expressed [Source:HGNC Symbol;Acc:27338] 66,657 4ENSG00000204991 Q8WWL2 SPIRE2 spire homolog 2 (Drosophila) [Source:HGNC Symbol;Acc:30623] 79,671 3;4
TARGET USING GLOBAL MASS LIST
LC CONDITIONSGradient Profile 100 min from 0 to 35% B
MS SYSTEMAcquisition method Global mass list
Cell Line Fraction Peptide Protein Score PEP
CCD18 2 EFQEEMAALK Q8TBY8; Q8TBY8-2; Q8TBY8-3 66.19 0.023021
CCD18 2 LMEGELEALR Q8TBY8; Q8TBY8-2 66.435 0.023902
MAXQUANT
Proteotypic peptides obtained at SRM Atlas
Database: uniprot_sprot_human_HPP
Q8TBY8 isoforms alignment
Q8TBY8 isoforms 1 and 2 are present in both CCD18 and Ramos cell lines
