Proteomics

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Proteomic analysis of separated Golgi cisternae gives insights into secretory pathway organization Proteomic analysis of separated Golgi cisternae gives insights into secretory pathway organization

ABSTRACT: The Golgi apparatus is the hub of the secretory pathway, trafficking proteins and lipids and synthesizing complex glycans. The spatial distribution of resident proteins across individual Golgi cisternae is functionally essential for complex molecule biosynthesis. However, the inability to-date to separate Golgi cisternae means both the distribution of most residents, and the mechanisms determining their distribution, are unknown. Here, we exploit differences in surface charge between membranes to perform the first separation of Golgi cisternae. We localize over 400 proteins to the Golgi and describe the cisternal distribution of over 250 Golgi residents, as well as complex glycans. Both protein and glycan distributions are validated in-vivo using super-resolution microscopy. Results reveal distinct functional compartmentalization amongst resident Golgi proteins, which relates to the distribution of carbohydrate reaction products. Analysis of cisternal proteomes shows that exoplasmic protein pI, exoplasmic hydrophobicity, Ser content and asymmetric Phe distribution increase across the Golgi.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Cell Suspension Culture

SUBMITTER: Harriet Parsons  

LAB HEAD: Harriet T Parsons

PROVIDER: PXD009978 | Pride | 2019-09-11

REPOSITORIES: Pride

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Publications

The order of enzymatic activity across Golgi cisternae is essential for complex molecule biosynthesis. However, an inability to separate Golgi cisternae has meant that the cisternal distribution of most resident proteins, and their underlying localization mechanisms, are unknown. Here, we exploit differences in surface charge of intact cisternae to perform separation of early to late Golgi subcompartments. We determine protein and glycan abundance profiles across the Golgi; over 390 resident pro  ...[more]

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