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

Q8TBY8 ISOFORMS ALIGNMENT

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

EFQEEMAALK

LMEGELEALR

Q8TBY8 isoforms alignment

Q8TBY8 isoforms 1 and 2 are present in both CCD18 and Ramos cell lines