PXD031097

PXD031097 is an original dataset announced via ProteomeXchange.

Title	Oligomerization, trans-reduction, and instability of mutant NOTCH3 in inherited vascular dementia
Description	Cerebral small vessel disease (SVD) is a prevalent disease of aging and a major contributor to stroke and dementia. The most commonly inherited SVD, CADASIL, is caused by dominantly acting cysteine-altering mutations in NOTCH3. These mutations change the number of cysteines from an even to an odd number, but the impact of these alterations on NOTCH3 protein structure remain unclear. Here, we prepared wildtype and four mutant recombinant NOTCH3 protein fragments to analyze the impact of CADASIL mutations on oligomerization, thiol status, and protein stability. Using gel electrophoresis, tandem MS/MS, and collision-induced unfolding, we find that NOTCH3 mutant proteins feature increased amounts of inappropriate disulfide bridges, reduced cysteines, and structural instability. Presence of a second protein factor, an N-terminal fragment of NOTCH3 (NTF), is capable of further altering disulfide statuses of both wildtype and mutant proteins, leading to increased numbers of reduced cysteines and further destabilization of NOTCH3 structure. In sum, these studies identify specific cysteine residues alterations and quaternary structure induced by CADASIL mutations in NOTCH3; further, we validate that reductive factors alter the structure and stability of this small vessel disease protein.
HostingRepository	PRIDE
AnnounceDate	2022-05-20
AnnouncementXML	Submission_2022-05-20_07:48:33.371.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kelly Young
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	N-ethylmaleimide derivatized cysteine; monohydroxylated residue; iodoacetamide derivatized residue; deamidated residue
Instrument	Q Exactive HF; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-19 09:37:08	ID requested
⏵ 1	2022-05-20 07:48:35	announced

Young KZ, Rojas Ram, í, rez C, Keep SG, Gatti JR, Lee SJ, Zhang X, Ivanova MI, Ruotolo BT, Wang MM, Oligomerization, trans-reduction, and instability of mutant NOTCH3 in inherited vascular dementia. Commun Biol, 5(1):331(2022) [pubmed]

submitter keyword: CADASIL, NOTCH3, disulfides

Michael M. Wang
contact affiliation	Departments of Neurology, University of Michigan, Ann Arbor, MI 48109-5622, USA. Molecular & Integrative Physiology, University of Michigan, Ann Arbor, MI 48109-5622, USA Neurology Service, VA Ann Arbor Healthcare System, Ann Arbor, MI 48105, USA
contact email	micwang@umich.edu
lab head
Kelly Young
contact affiliation	University of Michigan
contact email	kzyoung@umich.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD031097
     1. Label: PRIDE project
     2. Name: Oligomerization, trans-reduction, and instability of mutant NOTCH3 in inherited vascular dementia