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Please include the following statement when referencing the CPTAC Assay Portal
We would like to acknowledge the National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (CPTAC) Assay Portal (assays.cancer.gov) for developing assays and establishing criteria for the assays described in this publication.

Overview Data source: UniProt

Official Gene Symbol Other Aliases
RPS6KA1 MAPKAPK1A, RSK1
Sequence Length (AA) Molecular Weight (Da)
735 82723
Protein Name
Ribosomal protein S6 kinase alpha-1
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MPLAQLKEPW PLMELVPLDP ENGQTSGEEA GLQPSKDEGV LKEISITHHV
60 70 80 90 100
KAGSEKADPS HFELLKVLGQ GSFGKVFLVR KVTRPDSGHL YAMKVLKKAT
110 120 130 140 150
LKVRDRVRTK MERDILADVN HPFVVKLHYA FQTEGKLYLI LDFLRGGDLF
160 170 180 190 200
TRLSKEVMFT EEDVKFYLAE LALGLDHLHS LGIIYRDLKP ENILLDEEGH
210 220 230 240 250
IKLTDFGLSK EAIDHEKKAY SFCGTVEYMA PEVVNRQGHS HSADWWSYGV
260 270 280 290 300
LMFEMLTGSL PFQGKDRKET MTLILKAKLG MPQFLSTEAQ SLLRALFKRN
310 320 330 340 350
PANRLGSGPD GAEEIKRHVF YSTIDWNKLY RREIKPPFKP AVAQPDDTFY
360 370 380 390 400
FDTEFTSRTP KDSPGIPPSA GAHQLFRGFS FVATGLMEDD GKPRAPQAPL
410 420 430 440 450
HSVVQQLHGK NLVFSDGYVV KETIGVGSYS ECKRCVHKAT NMEYAVKVID
460 470 480 490 500
KSKRDPSEEI EILLRYGQHP NIITLKDVYD DGKHVYLVTE LMRGGELLDK
510 520 530 540 550
ILRQKFFSER EASFVLHTIG KTVEYLHSQG VVHRDLKPSN ILYVDESGNP
560 570 580 590 600
ECLRICDFGF AKQLRAENGL LMTPCYTANF VAPEVLKRQG YDEGCDIWSL
610 620 630 640 650
GILLYTMLAG YTPFANGPSD TPEEILTRIG SGKFTLSGGN WNTVSETAKD
660 670 680 690 700
LVSKMLHVDP HQRLTAKQVL QHPWVTQKDK LPQSQLSHQD LQLVKGAMAA
710 720 730 735
TYSALNSSKP TPQLKPIESS ILAQRRVRKL PSTTL

Data source: UniProt


Position of Targeted Peptide Analytes Relative to SNPs, Isoforms, and PTMs

Uniprot Database Entry PhosphoSitePlus ®

Click a point on a node
to view detailed assay information below
All other points link out to UniProt



Phosphorylation Acetylation Ubiquitylation Other

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Assay Details for non-CPTAC-5446 Collapse assay details

Data source: Panorama

Official Gene Symbol
RPS6KA1
Peptide Sequence
GFSFVATGLMEDDGKPR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
378
Peptide End
394
CPTAC ID
non-CPTAC-5446
Peptide Molecular Mass
1,825.8669
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y8 (1+) 11.8 5.3 3.7 11.5 5.5 3.8 16.5 7.6 5.3 15 15 15
y12 (2+) 7.5 4.4 1.5 9.6 6.2 2.9 12.2 7.6 3.3 15 15 15
y13 (2+) 11.4 4.1 2.5 7.1 6.3 4 13.4 7.5 4.7 15 15 15
y15 (2+) 4.2 2.9 2.9 6.1 4.4 2.9 7.4 5.3 4.1 15 15 15
sum 3.4 3 1.8 3.9 5.1 2.1 5.2 5.9 2.8 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5702 Collapse assay details

Data source: Panorama

Official Gene Symbol
RPS6KA1
Peptide Modified Sequence
GFS[+79.966331]FVATGLMEDDGKPR
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
378
Peptide End
394
CPTAC ID
non-CPTAC-5702
Peptide Molecular Mass
1,905.8332
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y4 (1+) 15.8 6.7 3.4 15.7 7.3 4.6 22.3 9.9 5.7 15 15 15
y10 (1+) 12 8.2 8.7 13.7 7.4 10.6 18.2 11 13.7 15 15 15
b4-98 (1+) 17.4 7.6 9.4 21.7 10 10.1 27.8 12.6 13.8 15 15 15
y11 (1+) 14.6 8.5 9.6 14.3 9.9 8.5 20.4 13 12.8 15 15 15
y12 (1+) 12 4.8 4 9.8 5.8 5 15.5 7.5 6.4 15 15 15
sum 7.6 3.8 3.5 8.1 4.6 3.5 11.1 6 4.9 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-2995 Collapse assay details

Data source: Panorama

Official Gene Symbol
RPS6KA1
Peptide Modified Sequence
ETIGVGSYSEC[+57.0]K
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
433
Peptide - Site of Modification
11
Peptide Start
422
Peptide End
433
CPTAC ID
CPTAC-2995
Peptide Molecular Mass
1,328.5918
Species
Homo Sapiens
Assay Type
Direct MRM
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y7 (1+) 5.9 4.4 3.1 11.5 5.3 5.4 12.9 6.9 6.2 15 15 15
y9 (1+) 6.3 6.1 2.1 11.6 6.1 4.9 13.2 8.6 5.3 15 15 15
y4 (1+) 4.5 6.8 3.2 8.8 4.8 5 9.9 8.3 5.9 15 15 15
sum 5.4 5 2.5 11 5.4 5 12.3 7.4 5.6 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-2994 Collapse assay details

Data source: Panorama

Official Gene Symbol
RPS6KA1
Peptide Sequence
NLVFSDGYVVK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
411
Peptide End
421
CPTAC ID
CPTAC-2994
Peptide Molecular Mass
1,239.6499
Species
Homo Sapiens
Assay Type
Direct MRM
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y8 (1+) 4.5 5.2 4.2 11.6 7.1 6 12.4 8.8 7.3 15 15 15
y9 (1+) 9 4.2 5.2 16.3 7.5 6.7 18.6 8.6 8.5 15 15 15
b3 (1+) 3.4 4 3.5 9.6 4.9 5.2 10.2 6.3 6.3 15 15 15
sum 4.4 4.3 3.8 11.7 6.2 5.5 12.5 7.5 6.7 15 15 15


Additional Resources and Comments