TY - JOUR
T1 - Fast and deep phosphoproteome analysis with the Orbitrap Astral mass spectrometer
AU - Lancaster, Noah M.
AU - Sinitcyn, Pavel
AU - Forny, Patrick
AU - Peters-Clarke, Trenton M.
AU - Fecher, Caroline
AU - Smith, Andrew J.
AU - Shishkova, Evgenia
AU - Arrey, Tabiwang N.
AU - Pashkova, Anna
AU - Robinson, Margaret Lea
AU - Arp, Nicholas
AU - Fan, Jing
AU - Hansen, Juli
AU - Galmozzi, Andrea
AU - Serrano, Lia R.
AU - Rojas, Julie
AU - Gasch, Audrey P.
AU - Westphall, Michael S.
AU - Stewart, Hamish
AU - Hock, Christian
AU - Damoc, Eugen
AU - Pagliarini, David J.
AU - Zabrouskov, Vlad
AU - Coon, Joshua J.
N1 - Publisher Copyright:
© The Author(s) 2024.
PY - 2024/12
Y1 - 2024/12
N2 - Owing to its roles in cellular signal transduction, protein phosphorylation plays critical roles in myriad cell processes. That said, detecting and quantifying protein phosphorylation has remained a challenge. We describe the use of a novel mass spectrometer (Orbitrap Astral) coupled with data-independent acquisition (DIA) to achieve rapid and deep analysis of human and mouse phosphoproteomes. With this method, we map approximately 30,000 unique human phosphorylation sites within a half-hour of data collection. The technology is benchmarked to other state-of-the-art MS platforms using both synthetic peptide standards and with EGF-stimulated HeLa cells. We apply this approach to generate a phosphoproteome multi-tissue atlas of the mouse. Altogether, we detect 81,120 unique phosphorylation sites within 12 hours of measurement. With this unique dataset, we examine the sequence, structural, and kinase specificity context of protein phosphorylation. Finally, we highlight the discovery potential of this resource with multiple examples of phosphorylation events relevant to mitochondrial and brain biology.
AB - Owing to its roles in cellular signal transduction, protein phosphorylation plays critical roles in myriad cell processes. That said, detecting and quantifying protein phosphorylation has remained a challenge. We describe the use of a novel mass spectrometer (Orbitrap Astral) coupled with data-independent acquisition (DIA) to achieve rapid and deep analysis of human and mouse phosphoproteomes. With this method, we map approximately 30,000 unique human phosphorylation sites within a half-hour of data collection. The technology is benchmarked to other state-of-the-art MS platforms using both synthetic peptide standards and with EGF-stimulated HeLa cells. We apply this approach to generate a phosphoproteome multi-tissue atlas of the mouse. Altogether, we detect 81,120 unique phosphorylation sites within 12 hours of measurement. With this unique dataset, we examine the sequence, structural, and kinase specificity context of protein phosphorylation. Finally, we highlight the discovery potential of this resource with multiple examples of phosphorylation events relevant to mitochondrial and brain biology.
UR - http://www.scopus.com/inward/record.url?scp=85201389924&partnerID=8YFLogxK
U2 - 10.1038/s41467-024-51274-0
DO - 10.1038/s41467-024-51274-0
M3 - Article
C2 - 39147754
AN - SCOPUS:85201389924
SN - 2041-1723
VL - 15
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 7016
ER -