Dataset Information

Systematic Discovery of Deeply Conserved Plant Protein Complexes: hemp

ABSTRACT: The associated files are mass spec data from mixed-bed ion exchange chromatography fractions. The starting material wasa clarified homogenate of Trader Joe's "Raw Shelled Hemp Seed".

INSTRUMENT(S):

ORGANISM(S): Cannabis Sativa

TISSUE(S): Embryo

SUBMITTER: Ophelia Papoulas

LAB HEAD: Edward M. Marcotte

PROVIDER: PXD012969 | Pride | 2019-10-28

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Hempseed_IEX_18a_07202017..comet.pep.xml	Pepxml
	Hempseed_IEX_18a_07202017.MSGF+.pride.mztab.gz	Mztab
	Hempseed_IEX_18a_07202017.MSGF.mzid.gz	Mzid
	Hempseed_IEX_18a_07202017.mzXML	Mzxml
	Hempseed_IEX_18a_07202017.raw	Raw

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Project description:Background and Aims Aging is the primary risk factor for cardiovascular disease (CVD), but the mechanisms underlying age-linked atherosclerosis remain unclear. We previously observed that long-lived vascular matrix proteins can acquire ‘gain-of-function’ isoDGR motifs that might play a role in atherosclerotic pathology. Methods: IsoDGR-specific mAb were generated and used for ELISA-based measurement of motif levels in plasma samples from patients with coronary artery diseases (CAD) and non-CAD controls. Functional consequences of isoDGR accumulation in age-damaged fibronectin were determined by bioassay for capacity to activate monocytes, macrophages, and endothelial cells (signalling activity, pro-inflammatory cytokine expression, and recruitment/adhesion potential). Mice deficient in the isoDGR repair enzyme PCMT1 were used to assess motif distribution and macrophage localisation in vivo. Results: IsoDGR-modified fibronectin and fibrinogen levels in patient plasma were significantly enhanced in CAD and further associated with smoking status. Functional assays demonstrated that isoDGR-modified fibronectin activated both monocytes and macrophages via integrin receptor ‘outside in’ signalling, triggering an ERK:AP-1 cascade and expression of pro-inflammatory cytokines MCP-1 and TNF to drive additional recruitment of circulating leukocytes. IsoDGR-modified fibronectin also induced endothelial cell expression of integrin 1 to further enhance cellular adhesion and matrix deposition. Analysis of murine aortic tissues confirmed accumulation of isoDGR-modified proteins co-localised with CD68+ macrophages in vivo. Conclusions: Age-damaged fibronectin features isoDGR motifs that increase binding to integrins on the surface of monocytes, macrophages, and endothelial cells. Subsequent activation of ‘outside-in’ signalling elicits a range of potent cytokines and chemokines that drive additional leukocyte recruitment to the developing atherosclerotic matrix.

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Systematic Discovery of Deeply Conserved Plant Protein Complexes: hemp

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