CAGE-TRX expands the scope of time-resolved crystallography through genetically encoded active-site photocaging

Year: 2026

Type: preprint

Abstract: We present CAGE-TRX, a broadly applicable time-resolved strategy for release-quench-probe cryocrystallography and release-probe room temperature serial crystallography. These workflows enable light-triggered control of enzyme activity via genetically encoded photocaged amino acids. By decoupling reaction initiation from substrate design, this approach allows synchronized catalysis in crystallo and the capture of transient intermediates. Using β-lactamases as model systems, we demonstrate efficient decaging, restoration of activity, and structural visualization of reaction intermediates.

Source: bioRxiv (Cold Spring Harbor Laboratory)

DOI: 10.64898/2026.06.26.734858

Authors: Clyde A. Smith, Ailiena O. Maggiolo, Chasity Janosko, Maura E. Charette, Nilakshi Paul, Marta Toth, Guillermo Calero, Sierra M. Carr, Silvia Russi, Sergei B. Vakulenko, Alexander Deiters, Aina E. Cohen

Institutions: Stanford Synchrotron Radiation Lightsource, Stanford University, University of Pittsburgh, University of Notre Dame

Language: English

Topic: Enzyme Structure and Function

Subfield: Materials Chemistry

Field: Materials Science

Domain: Physical Sciences

Open Access status: green

Funders: U.S. Department of Energy, National Institutes of Health, Office of Science, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, Basic Energy Sciences, Biological and Environmental Research, Laboratory Directed Research and Development

OpenAlex ID: W7166530653