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Identification of potential self-interaction sites on golgin Imh1-C213 by cross-linking mass spectrometry

ABSTRACT: The golgins are long coiled-coil proteins involved in vesicular transport to the Golgi, a process that contributes to Golgi function and integrity. Previous studies have elucidated that their self-interaction and their interaction with small GTPase Arl1 are critical for their Golgi localization. Here, we report that glycerol regulates Golgi localization of Imh1, a prototypic yeast golgin. Experimentally, we observed that cellular conditions leading to reduced intracellular glycerol levels trigger the release of Imh1 from the Golgi, whereas restoring glycerol levels reverses this effect. Given that glycerol is a well-documented molecular chaperone, we investigated its impact on the conformation of the Imh1 C-terminal fragment (Imh1-C213) in vitro, employing limited proteolysis assays. To further understand glycerol-induced conformational changes, we used isotope-labeled BS3-d0/d4 crosslinkers in combination with LC-MS/MS to map potential self-interaction sites on Imh1-C213. Our findings reveal and quantify several distinct cross-linked peptide pairs within Imh1-C213, in both the presence and absence of glycerol. These observed differences likely correspond to glycerol-induced conformational changes, which provide novel insights into the structural dynamics of golgins and their functional regulation within the Golgi apparatus.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Chia-Jung Yu

LAB HEAD: Chia-Jung Yu

PROVIDER: PXD061125 | Pride | 2025-04-30

REPOSITORIES: Pride

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	1-0400-0800.raw	Raw
	1-0700-1100.raw	Raw
	1-1000-1400.raw	Raw
	1SW-0400-0800.raw	Raw
	1SW-0700-1100.raw	Raw

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