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BS3 crosslinking of the Anaphase-Promoting Complex/Cyclosome (APC/C)

ABSTRACT: How Apc8-LOOP affects the displacement of Apc1-300LOOP was investigated using crosslinking mass spectrometry (MS) to gain insight into the point of interaction between disordered Apc8-LOOP and the APC/C center structure.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Xenopus Laevis (african Clawed Frog)

SUBMITTER: Juan Zou

LAB HEAD: Juri Rappsilber

PROVIDER: PXD044827 | Pride | 2024-09-08

REPOSITORIES: Pride

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			Action	DRS
	XenopusAPCC.fasta	Fasta
	Zou_Rappsilber_HY_APCC_BS3_F1.mgf	Mgf
	Zou_Rappsilber_HY_APCC_BS3_F1.raw	Raw
	Zou_Rappsilber_HY_APCC_BS3_F2.mgf	Mgf
	Zou_Rappsilber_HY_APCC_BS3_F2.raw	Raw

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BS3 crosslinking of the Anaphase-Promoting Complex/Cyclosome (APC/C)

Project description:In order to investigate the dynamic regulation of the APC/C by intrinsically disordered regions (IDRs) upon phosphorylation, we employed cross-linking mass spectrometry (CLMS). Given that the phosphorylation of Apc1-300L (IDR in Apc1) is a key determinant in its release from the co-activator binding site, we compared the interaction profiles of unphosphorylated and hyper-phosphorylated rAPC/C_WT complexes. The CLMS analysis highlights not only known interactions consistently identified within APC/C complexes, regardless of phosphorylation status, but also dynamic IDR-mediated interactions that are lost upon phosphorylation or are phosphorylation-dependent. Additionally, the data indicate that phosphorylation significantly reduces Apc1-300L's interaction with Apc8-L (IDR in Apc8) and Apc6, suggesting Apc1-300L's dislocation from the co-activator Cdc20 binding site, thereby facilitating Cdc20 loading onto the APC/C.

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