PXD010258

PXD010258 is an original dataset announced via ProteomeXchange.

Title	The Plethora of Angiotensin-Converting Enzyme Substrates in Mouse Plasma
Description	Angiotensin converting enzyme (ACE) is known for converting inactive angiotensin I into the potent vasoconstrictor angiotensin II, playing a critical role in blood pressure regulation. However, there is evidence that the dicarboxypeptidase activity of ACE is also essential for other physiological processes likely through processing of various peptide substrates. This study used mass spectrometry for the comprehensive detection and identification of natural substrates and products of ACE within the mouse plasma peptidome. The plasma peptidome of ACE KO mice was obtained through a multi-step purification process which included organic TCA precipitation as well as size exclusion and reversed phase chromatography. The obtained complex mixture of endogenous peptides was then subjected to in vitro cleavage by ACE. ACE-treated and untreated samples were then analyzed by LC/MS on an Orbitrap mass spectrometer, followed by alignment of MS1 data by Progenesis QI software. The MS1 signals that gained or lost intensity after treatment with ACE, were considered as possible products and substrates of ACE, respectively, and were selected for a targeted MS/MS analysis, and subsequently identified with PEAKS 8.5 and Proteome Discoverer software. Results. Close to 250 natural peptides were identified as possible substrates and products of ACE, demonstrating the high promiscuity of the enzyme. The use of internal standards as well as detection of some expected endogenous peptides, such as angiotensin II and bradykinin, supported the validity of the approach. Some of the newly identified substrates of ACE are known for their biological activities. For example, a fragment of complement C3, the 17-amino acid peptide C3f, exhibits spasminogenic activity and was processed by ACE. ACE cleavage of select peptides was further confirmed in vitro. Also, concentrations of ACE substrates in plasma from mice with variant genetic ACE domain backgrounds were determined by LC/MS using multiple reaction monitoring on a triple quadrupole mass spectrometer. The in vivo results were consistent with the in vitro results, in the sense that higher levels of the ACE substrates were observed when the respective processing domain was knocked out. The use of transgenic mice as well as ACE with single active domain allowed clarifying the ACE domain selectivity towards individual peptide substrates. This study resulted in creation of a library of substrates and products of ACE that can be further tested for their biological function and can help to elucidate the link between ACE and the numerous physiological effects attributed to its activity.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:46:02.715.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD010258
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Margarita (Rita) Semis
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	Oxidation; Carbamidomethyl
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2018-06-28 01:44:34	ID requested
1	2019-11-12 11:43:39	announced
⏵ 2	2024-10-22 04:46:08	announced	2024-10-22: Updated project metadata.

10.6019/PXD010258;

Semis M, Gugiu GB, Bernstein EA, Bernstein KE, Kalkum M, The Plethora of Angiotensin-Converting Enzyme-Processed Peptides in Mouse Plasma. Anal Chem, 91(10):6440-6453(2019) [pubmed]

10.1021/acs.analchem.8b03828;

curator keyword: Biological, Biomedical

submitter keyword: Peptidomics

Markus Kalkum
contact affiliation	Department of Molecular Imaging and Therapy, Diabetes and Metabolism Research Institute, Beckman Research Institute of the City of Hope, USA Mass Spectrometry & Proteomics Core Facility, Beckman Research Institute of the City of Hope, USA
contact email	mkalkum@coh.org
lab head
Margarita (Rita) Semis
contact affiliation	City of Hope Medical Center, Beckman Research Institute, Department of Molecular Imaging and Therapy
contact email	msemis@coh.org
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD010258
     1. Label: PRIDE project
     2. Name: The Plethora of Angiotensin-Converting Enzyme Substrates in Mouse Plasma