6LTV

Crystal Structure of I122A/I330A variant of S-adenosylmethionine synthetase from Cryptosporidium hominis in complex with ONB-SAM (2-nitro benzyme S-adenosyl-methionine)

Summary for 6LTV

• Entry DOI: 10.2210/pdb6ltv/pdb
• Descriptor: S-adenosylmethionine synthase, TRIPHOSPHATE, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: triphosphate, s-adenosylmethionine synthetase, transferase
• Biological source: Cryptosporidium hominis
• Total number of polymer chains: 2
• Total formula weight: 93347.93

Authors

Singh, R.K.,Michailidou, F.,Rentmeister, A.,Kuemmel, D. (deposition date: 2020-01-23, release date: 2020-10-21, Last modification date: 2023-11-29)

Primary citation

Michailidou, F.,Klocker, N.,Cornelissen, N.V.,Singh, R.K.,Peters, A.,Ovcharenko, A.,Kummel, D.,Rentmeister, A.
Engineered SAM Synthetases for Enzymatic Generation of AdoMet Analogs with Photocaging Groups and Reversible DNA Modification in Cascade Reactions.
Angew.Chem.Int.Ed.Engl., 60:480-485, 2021

PubMed Abstract: Methylation and demethylation of DNA, RNA and proteins has emerged as a major regulatory mechanism. Studying the function of these modifications would benefit from tools for their site-specific inhibition and timed removal. S-Adenosyl-L-methionine (AdoMet) analogs in combination with methyltransferases (MTases) have proven useful to map or block and release MTase target sites, however their enzymatic generation has been limited to aliphatic groups at the sulfur atom. We engineered a SAM synthetase from Cryptosporidium hominis (PC-ChMAT) for efficient generation of AdoMet analogs with photocaging groups that are not accepted by any WT MAT reported to date. The crystal structure of PC-ChMAT at 1.87 Å revealed how the photocaged AdoMet analog is accommodated and guided engineering of a thermostable MAT from Methanocaldococcus jannaschii. PC-MATs were compatible with DNA- and RNA-MTases, enabling sequence-specific modification ("writing") of plasmid DNA and light-triggered removal ("erasing").

PubMed: 33017502
DOI: 10.1002/anie.202012623

Experimental method

X-RAY DIFFRACTION (1.87 Å)