8D6H

Q108K:K40L:T51C:T53A:R58L:Q38F:Q4F mutant of hCRBPII bound to synthetic fluorophore CM1V after UV irradiation

Summary for 8D6H

• Entry DOI: 10.2210/pdb8d6h/pdb
• Descriptor: Retinol-binding protein 2, GLYCEROL, ACETATE ION, ... (5 entities in total)
• Functional Keywords: transport protein, hcrbpii
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 16996.04

Authors

Bingham, C.R.,Geiger, J.H.,Borhan, B. (deposition date: 2022-06-06, release date: 2023-02-01, Last modification date: 2024-11-13)

Primary citation

Maity, S.,Bingham, C.,Sheng, W.,Ehyaei, N.,Chakraborty, D.,Tahmasebi-Nick, S.,Kimmel, T.E.,Vasileiou, C.,Geiger, J.H.,Borhan, B.
Light controlled reversible Michael addition of cysteine: a new tool for dynamic site-specific labeling of proteins.
Analyst, 148:1085-1092, 2023

PubMed Abstract: Cysteine-based Michael addition is a widely employed strategy for covalent conjugation of proteins, peptides, and drugs. The covalent reaction is irreversible in most cases, leading to a lack of control over the process. Utilizing spectroscopic analyses along with X-ray crystallographic studies, we demonstrate Michael addition of an engineered cysteine residue in human Cellular Retinol Binding Protein II (hCRBPII) with a coumarin analog that creates a non-fluorescent complex. UV-illumination reverses the conjugation, yielding a fluorescent species, presumably through a -Michael process. This series of events can be repeated between a bound and non-bound form of the cysteine reversibly, resulting in the ON-OFF control of fluorescence. The details of the mechanism of photoswitching was illuminated by recapitulation of the process in light irradiated single crystals, confirming the mechanism at atomic resolution.

PubMed: 36722993
DOI: 10.1039/d2an01395a

Experimental method

X-RAY DIFFRACTION (1.6 Å)