S-adenosyl-1-methionine(SAM)-dependent enzymes regulate various disease-related behaviors in all organisms.Recently,the leporin biosynthesis enzyme LepI,a SAM-dependent enzyme,was reported to catalyze pericyclic react...S-adenosyl-1-methionine(SAM)-dependent enzymes regulate various disease-related behaviors in all organisms.Recently,the leporin biosynthesis enzyme LepI,a SAM-dependent enzyme,was reported to catalyze pericyclic reactions in leporin biosynthesis;however,the mechanisms underlying LepI activation and catalysis remain unclear.This study aimed to investigate the molecular mechanisms of LepI.Here,we reported crystal structures of LepI bound to SAM/5′-deoxy-5′-(methylthio)adenosine(MTA),S-adenosyl-homocysteine(SAH),and SAM/substrate states.Structural and biochemical analysis revealed that MTA or SAH inhibited the enzyme activities,whereas SAM activated the enzyme.The analysis of the substrate-bound structure of LepI demonstrated that this enzymatic retro-Claisen rearrangement was primarily driven by three critical polar residues His133,Arg197,Arg295 around the active site and assisted by SAM with unclear mechanism.The present studies indicate that the unique mechanisms underlying regulatory and catalysis of the unusual SAM-dependent enzyme LepI,not only strengthening current understanding of the fundamentally biochemical catalysis,but also providing novel insights into the design of SAM-dependent enzyme-specific small molecules.展开更多
基金We gratefully acknowledge the financial support from the National Natural Science Foundation of China(Grant No.31570842,No.31870836,and No.21702141)the National Young Thousand Talents Program and the Sichuan Province Thousand Talents program in China.
文摘S-adenosyl-1-methionine(SAM)-dependent enzymes regulate various disease-related behaviors in all organisms.Recently,the leporin biosynthesis enzyme LepI,a SAM-dependent enzyme,was reported to catalyze pericyclic reactions in leporin biosynthesis;however,the mechanisms underlying LepI activation and catalysis remain unclear.This study aimed to investigate the molecular mechanisms of LepI.Here,we reported crystal structures of LepI bound to SAM/5′-deoxy-5′-(methylthio)adenosine(MTA),S-adenosyl-homocysteine(SAH),and SAM/substrate states.Structural and biochemical analysis revealed that MTA or SAH inhibited the enzyme activities,whereas SAM activated the enzyme.The analysis of the substrate-bound structure of LepI demonstrated that this enzymatic retro-Claisen rearrangement was primarily driven by three critical polar residues His133,Arg197,Arg295 around the active site and assisted by SAM with unclear mechanism.The present studies indicate that the unique mechanisms underlying regulatory and catalysis of the unusual SAM-dependent enzyme LepI,not only strengthening current understanding of the fundamentally biochemical catalysis,but also providing novel insights into the design of SAM-dependent enzyme-specific small molecules.
