Dataset Information

Post-translational modifications of vaccine candidates of Cryptosporidium parvum

ABSTRACT: Here we describe the post translational modifications on vaccine candidate proteins from Cryptosporidium parvum. We have identified O-glycosylated peptides from GP15, GP40, GP900, and a novel mucin-like protein referred to as GP20. In addition, we have discovered that the immune dominant antigen, CP23, contains both N-myristoyl and C-palmitoyl modifications on the N-terminal region of the protein.

INSTRUMENT(S): Q Exactive Plus, LTQ Orbitrap XL ETD

ORGANISM(S): Cryptosporidium Parvum

TISSUE(S): Sporulated Oocyst

SUBMITTER: John Haserick

LAB HEAD: Catherine E. Costello

PROVIDER: PXD005989 | Pride | 2017-08-14

REPOSITORIES: Pride

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Publications

Cryptosporidium parvum vaccine candidates are incompletely modified with O-linked-N-acetylgalactosamine or contain N-terminal N-myristate and S-palmitate.

Haserick John R JR Klein Joshua A JA Costello Catherine E CE Samuelson John J

PloS one 20170808 8

Cryptosporidium parvum (studied here) and Cryptosporidium hominis are important causes of diarrhea in infants and immunosuppressed persons. C. parvum vaccine candidates, which are on the surface of sporozoites, include glycoproteins with Ser- and Thr-rich domains (Gp15, Gp40, and Gp900) and a low complexity, acidic protein (Cp23). Here we used mass spectrometry to determine that O-linked GalNAc is present in dense arrays on a glycopeptide with consecutive Ser derived from Gp40 and on glycopeptid ...[more]

PMID: 28792526

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Project 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.

Dataset Information

Post-translational modifications of vaccine candidates of Cryptosporidium parvum

Publications

Cryptosporidium parvum vaccine candidates are incompletely modified with O-linked-N-acetylgalactosamine or contain N-terminal N-myristate and S-palmitate.

Similar Datasets

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