title: Protein splicing: its discovery and structural insight into novel chemical mechanisms.
abstract: Protein splicing is a posttranslational cellular process, in which an intervening protein sequence (intein) is self-catalytically excised out from a nascent protein precursor and the two flanking sequences (N- and C-exteins) are ligated to produce two mature enzymes. This unique reaction was first discovered from studies of the structure and expression of the VMA1 gene in Saccharomyces cerevisiae. VMA1 consists of a single open reading frame and yet comprises two independent genetic information for Vma1p (a catalytic 70-kDa subunit of the vacuolar H+-ATPase) and VDE (a 50-kDa DNA endonuclease) as an in-frame spliced insert in the gene. Subsequent studies have demonstrated that protein splicing is not unique for the VMA1 precursor and there are many operons in nature, which implement genetic information editing at protein level. To elucidate its precise reaction mechanisms from a viewpoint of structure-directed chemistry, a series of crystal structural studies has been carried out with the use of splicing-inactive and slowly spliceable precursors of VMA1 recombinants. One precursor structure revealed that the N-terminal junction of the introduced extein polypeptide forms an intermediate containing a five-membered thiazolidine ring. The other precursor structures showed spliced products with a linkage between the N- and C-extein segments. This article summarizes biochemical and structural studies on a self-catalytic mechanism for protein splicing that is triggered and terminated solely via thiazolidine intermediates with tetrahedral configurations formed within the splicing sites where proton ingress and egress are driven by balanced protonation and deprotonation.
authors.affiliation: Department of Biosciences, Teikyo University of Science and Technology, Uenohara, Yamanashi, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Anraku
authors.forename: Ryuta
authors.initials: R
authors.lastname: Mizutani
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Satow
mesh_terms: D000595:Amino Acid Sequence; D003461:Crystallography; D047669:Exteins / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019154:Protein Splicing / Q000235:genetics* / Q000502:physiology*; D006180:Proton-Translocating ATPases / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013844:Thiazoles
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D013844:Thiazoles; D006180:Proton-Translocating ATPases; C081687:VMA1 protein, S cerevisiae