PXD018902

PXD018902 is an original dataset announced via ProteomeXchange.

Title	O-Fucosylation of ADAMTSL2 Thrombospondin Type 1 Repeats is Required for Secretion and is Impacted by Geleophysic Dysplasia-Causing Mutations
Description	A disintegrin-like and metalloprotease domain with thrombospondin type 1 repeats-like 2 (ADAMTSL2) is a matricellular protein that interacts with latent TGF-b binding protein 1 and fibrillin-1. ADAMTSL2 mutations cause an autosomal recessive connective tissue disorder named geleophysic dysplasia 1 (GPHYSD1, OMIM #231050), which is characterized by short stature, small hands and feet, and cardiac defects. ADAMTSL2 contains seven thrombospondin type-1 repeats (TSRs), six of which contain the consensus sequence for serine or threonine glycosylation by protein O-fucosyltransferase 2 (POFUT2). O-fucose modified TSRs are subsequently elongated to a Glucose1-3Fucose (GlcFuc) disaccharide by b3-glucosyltransferase (B3GLCT). B3GLCT mutations cause Peters Plus Syndrome (PTRPLS, OMIM #261540), which is characterized by skeletal defects similar to those of GPHYSD1. Several ADAMTSL2 TSRs have consensus sequences for C-mannosylation, and one TSR contains a sequon for N-glycosylation that overlaps with an O-fucosylation site. Six reported GPHYSD1 mutations occur within the TSRs and two lie near O-fucosylation sites. To investigate the effects of TSR glycosylation on ADAMTSL2 function, we used mass spectrometry to identify glycan modifications at the predicted consensus sequences. We found that most TSRs were modified with the GlcFuc disaccharide at high stoichiometry at O-fucosylation sites and variable mannose stoichiometry at C-mannosylation sites. Loss of ADAMTSL2 secretion in POFUT2 knockout but not in B3GLCT knockout cells suggested that impaired ADAMTSL2 secretion is not responsible for skeletal defects in PTRPLS patients lacking B3GLCT. In contrast, secretion of mouse ADAMTSL2 carrying GPHYSD1 mutations (S641L in TSR3 and G817R in TSR6) was significantly reduced in HEK293T cells. Moreover, S641L eliminated O-fucosylation of TSR3. Combined these results provide strong evidence that developmental abnormalities in GPHYSD1 patients with this mutation are caused by loss of O-fucosylation on TSR3 and impaired ADAMTSL2 secretion.
HostingRepository	PRIDE
AnnounceDate	2020-10-01
AnnouncementXML	Submission_2020-09-30_22:54:08.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ao Zhang
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-29 08:10:47	ID requested
⏵ 1	2020-09-30 22:54:08	announced

Zhang A, Berardinelli SJ, Leonhard-Melief C, Vasudevan D, Liu TW, Taibi A, Giannone S, Apte SS, Holdener BC, Haltiwanger RS, Fucosylation of ADAMTSL2 is required for secretion and is impacted by geleophysic dysplasia-causing mutations. J Biol Chem, 295(46):15742-15753(2020) [pubmed]

submitter keyword: Thrombospondin Type 1 Repeats, TSR glycosylation, ADAMTSL2 O-fucosylation, Geleophysic Dysplasia

Robert S. Haltiwanger
contact affiliation	Department of Biochemistry and Molecular Biology, Complex Carbohydrate Research Center, The University of Georgia, U.S.A.
contact email	rhalti@uga.edu
lab head
Ao Zhang
contact affiliation	The University of Georgia
contact email	az59150@uga.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD018902
     1. Label: PRIDE project
     2. Name: O-Fucosylation of ADAMTSL2 Thrombospondin Type 1 Repeats is Required for Secretion and is Impacted by Geleophysic Dysplasia-Causing Mutations