Dataset Information

SMAP3-ID for identification of protein-protein interactions in native cells

ABSTRACT: Proteins regulate biological functions through formation of distinct protein complexes. Identification and characterization of these protein-protein interactions are critical to decipher their mechanism of action. Different antibody- or crosslinking-based methods have been developed to identify the protein-protein interactions. However, these methods require genetic engineering and/or other means to disrupt the native environments. To circumvent this limitation, we introduce here SMAP3-ID (Small Molecule Assisted identification of Protein-Protein interactions through Proximity) method to identify protein-protein interactions in native cells. This method entails a selective ligand for binding to a protein of interest for photocrosslinking, a live-cell compatible bioorthogonal click reaction with a trifunctional chemical probe and a final photocrosslinking reaction to covalently capture the interacting proteins. Using SMAP3-ID method and nuclear lamin as an example, we identified lamin-interactors in native cells. Significantly, we identified a number of mitochondrial enzymes as novel lamin A (LA)-interactors. The interactions between mitochondrial enzymes and LA were further validated, which provides mechanistic insights underlying the metabolic alterations caused by mutations in LA. We conclude that SMAP3-ID is a powerful and generalizable method to identify protein-protein interactions in the native cellular environment.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Phillip Wilmarth

LAB HEAD: Dr. Xiangshu Xiao

PROVIDER: PXD054249 | Pride | 2025-05-25

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	2022.10_Homo-Sapiens_UP000005640_9606_both.fasta	Fasta
	MSConvert_GUI_log.txt	Txt
	PAW_protein_grouper.log	Other
	PAW_results.log	Other
	PAW_table_descriptions_9.txt	Txt

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Publications

SMAP3-ID for Identification of Endogenous Protein-Protein Interactions Reveals Regulation of Mitochondrial Activity by Lamins.

Warren Julia J Wang Jian J Dhoro Francis F Chao Bo B Reddy Ashok A Petrie Stefanie Kaech SK David Larry L LL Xiao Xiangshu X Li Bingbing X BX

JACS Au 20250114 1

Proteins regulate biological functions through the formation of distinct protein complexes. Identification and characterization of these protein-protein interactions are critical to deciphering their mechanism of action. Different antibody-based or cross-linking-based methods have been developed to identify the protein-protein interactions. However, these methods require genetic engineering or other means to disrupt the native environments. To circumvent this limitation, we introduce here SMAP3- ...[more]

PMID: 39886595

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