PXD030549

PXD030549 is an original dataset announced via ProteomeXchange.

Title	MLC phosphorylation by the kinase CaMKII
Description	MLC1 is membrane protein highly expressed by brain perivascular astrocytes. Mutations in MLC1 account for megalencephalic leukoencephalopathy with subcortical cysts (MLC), an incurable leu-kodystrophy characterized by macrocephaly, brain edema and cysts, myelin vacuolation and as-trocyte swelling, that cause cognitive and motor dysfunctions. MLC pathological models demon-strated that in astrocytes MLC1 mutations affect the swelling-activated Cl- currents (ICl,swell) medi-ated by volume-regulated anion channel (VRAC) and the consequent regulatory volume decrease (RVD), and also induce the abnormal activation of intracellular signaling pathways linked to in-flammation/osmotic stress. Despite this knowledge, MLC1 function and MLC molecular pathogen-esis are still elusive. Following the observations that calcium regulates all the MLC1-modulated processes and that intracellular Ca2+ homeostasis is altered in MLC1-defective cells, we applied biochemistry, molecular biology, video imaging, electrophysiology and proteomic techniques on cultured human and mouse astrocytes to study MLC1/calcium signaling relationships. We revealed that MLC1 binds the Ca2+ effector proteins calmodulin (CaM) and Ca2+/CaM-dependent protein ki-nase II (CaMKII) and, as result, changes its assembly, localization and functional properties in re-sponse to Ca2+ influx or release. Noteworthy, CaM binding to the COOH terminal promotes MLC1 trafficking to the plasma membrane, while CaMKII phosphorylation of the NH2 end potentiates MLC1 activation of ICl,swell, that are critically important for RVD. Overall, our results show that MLC1 is a Ca2+-regulated protein linking VRAC function and volume regulation to Ca2+ signaling in astro-cytes, opening new avenues to clarify the defective molecular pathways of MLC and other diseas-es where astrocyte swelling and brain edema underlie the pathological process. In this contest we submit here the MS data showing MLC phosphorylation on Threonine 17.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:32:38.523.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Serena Camerini
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	LTQ

Revision	Datetime	Status	ChangeLog Entry
0	2021-12-20 11:12:11	ID requested
1	2022-10-13 12:21:08	announced
⏵ 2	2023-11-14 08:32:39	announced	2023-11-14: Updated project metadata.

Brignone MS, Lanciotti A, Michelucci A, Mallozzi C, Camerini S, Catacuzzeno L, Sforna L, Caramia M, D'Adamo MC, Ceccarini M, Molinari P, Macioce P, Macchia G, Petrucci TC, Pessia M, Visentin S, Ambrosini E, -Dependence to Volume-Regulated Anion Channels (VRAC) in Astrocytes. Cells, 11(17):(2022) [pubmed]

submitter keyword: MLC Megalencephalic leukoencephalopathy phosphorylation mass spectrometry

Serena Camerini
contact affiliation	Italian National Institute of Health
contact email	serena.camerini@iss.it
lab head
Serena Camerini
contact affiliation	Istituto superiore di sanità
contact email	serena.camerini@iss.it
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/10/PXD030549

PRIDE project URI

• PRIDE
  1. PXD030549
     1. Label: PRIDE project
     2. Name: MLC phosphorylation by the kinase CaMKII