Light-activated BioID – an optically activated proximity labeling system to study protein–protein interactions DOI Creative Commons
Omer Shafraz, Carolyn Davis, Sanjeevi Sivasankar

и другие.

Journal of Cell Science, Год журнала: 2023, Номер 136(19)

Опубликована: Сен. 27, 2023

ABSTRACT Proximity labeling with genetically encoded enzymes is widely used to study protein–protein interactions in cells. However, the accuracy of proximity limited by a lack control over enzymatic process. Here, we present light-activated technology for mapping at cell membrane high and precision. Our technology, called BioID (LAB), fuses two halves split-TurboID enzyme photodimeric proteins CRY2 CIB1. We demonstrate, multiple lines, that upon illumination blue light, CIB1 dimerize, reconstitute initiate biotinylation. Turning off light leads dissociation halts benchmark LAB against TurboID method measuring proteome E-cadherin, an essential cell–cell adhesion protein. show can map E-cadherin-binding partners higher significantly fewer false positives than TurboID.

Язык: Английский

Photo-ANA enables profiling of host–bacteria protein interactions during infection DOI
Xiaomeng Li, Siyue Huang, Xiang Li

и другие.

Nature Chemical Biology, Год журнала: 2023, Номер 19(5), С. 614 - 623

Опубликована: Янв. 26, 2023

Язык: Английский

Процитировано

15
Depletion of endogenously biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in Caenorhabditis elegans DOI Creative Commons
Murat Artan, Markus Hartl, Wei‐Qiang Chen

и другие.

Journal of Biological Chemistry, Год журнала: 2022, Номер 298(9), С. 102343 - 102343

Опубликована: Авг. 3, 2022

Proximity-dependent protein labeling provides a powerful in vivo strategy to characterize the interactomes of specific proteins. We previously optimized proximity protocol for Caenorhabditis elegans using highly active biotin ligase TurboID. A significant constraint on sensitivity TurboID is presence abundant endogenously biotinylated proteins that take up bandwidth mass spectrometer, notably carboxylases use as cofactor. In C. elegans, these comprise POD-2/acetyl-CoA carboxylase alpha, PCCA-1/propionyl-CoA PYC-1/pyruvate carboxylase, and MCCC-1/methylcrotonyl-CoA alpha. Here, we developed ways remove prior streptavidin purification spectrometry by engineering their corresponding genes add C-terminal His

Язык: Английский

Процитировано

19
Creative approaches using proximity labeling to gain new biological insights DOI Creative Commons
Ryan R. Milione, Bin-Bin Schell, Cameron J. Douglas

и другие.

Trends in Biochemical Sciences, Год журнала: 2023, Номер 49(3), С. 224 - 235

Опубликована: Дек. 30, 2023

At its most fundamental level, life is a collection of synchronized cellular processes driven by interactions among biomolecules. Proximity labeling has emerged as powerful technique to capture these in native settings, revealing previously unexplored elements biology. This review highlights recent developments proximity labeling, focusing on methods that push the technologies beyond classic bait-prey paradigm, such RNA–protein interactions, ligand/small-molecule–protein cell surface protein and subcellular trafficking. The advancement address different biological problems will accelerate our understanding complex systems make up life.

Язык: Английский

Процитировано

13
Application of Mass Spectrometry for the Advancement of PROTACs DOI
Hao Yin,

Baoshuang Zhang,

Ruibing Chen

и другие.

Journal of Pharmaceutical and Biomedical Analysis, Год журнала: 2025, Номер unknown, С. 116829 - 116829

Опубликована: Март 1, 2025

Язык: Английский

Процитировано

0
Light-activated BioID – an optically activated proximity labeling system to study protein–protein interactions DOI Creative Commons
Omer Shafraz, Carolyn Davis, Sanjeevi Sivasankar

и другие.

Journal of Cell Science, Год журнала: 2023, Номер 136(19)

Опубликована: Сен. 27, 2023

ABSTRACT Proximity labeling with genetically encoded enzymes is widely used to study protein–protein interactions in cells. However, the accuracy of proximity limited by a lack control over enzymatic process. Here, we present light-activated technology for mapping at cell membrane high and precision. Our technology, called BioID (LAB), fuses two halves split-TurboID enzyme photodimeric proteins CRY2 CIB1. We demonstrate, multiple lines, that upon illumination blue light, CIB1 dimerize, reconstitute initiate biotinylation. Turning off light leads dissociation halts benchmark LAB against TurboID method measuring proteome E-cadherin, an essential cell–cell adhesion protein. show can map E-cadherin-binding partners higher significantly fewer false positives than TurboID.

Язык: Английский

Процитировано

10