Work

Structure of Ribosome-Inactivating Protein from Mirabilis jalapa and Its L12-Stalk-Dependent Inhibition of Escherichia coli Ribosome

Year: 2025

Type: article

Abstract: Mirabilis antiviral protein (MAP) is the type I ribosome-inactivating protein (RIP), which consists of an RNA N-glycosylase domain with no carbohydrate-binding domain. Unlike many RIPs, such as ricin or trichosanthin, which inactivate eukaryotic ribosomes, MAP also inactivates the E. coli ribosome by cleaving the N-glycosidic bond at A2660 of 23S ribosomal RNA. The structure of the wild-type MAP has not been revealed yet. Here, we expressed, purified, and crystallized the plural recombinant MAPs, including both E168Q and R171Q mutations (MAP-EQRQ) in E. coli, and determined the crystal structure of MAP-EQRQ at 2.1 Å resolution. According to the predicted structure with RNA (sarcin-ricin loop) and the mutant protein’s activities using quantitative RT-PCR, we showed that residue R171 at the active site of MAP is a key residue to form the stable complex with target adenine. Furthermore, we showed that MAP bound the C-terminal domains of eukaryotic P2-stalk as well as E. coli L12-stalk.

Source: Toxins

DOI: 10.3390/toxins17120575

Authors: Nanami Nishida, Yuki Ninomiya, Toru Yoshida, Takehito Tanzawa, Yasushi Maki, Hideji Yoshida, Hideaki Tsuge, Noriyuki Habuka

Institutions: Kyoto Sangyo University, Japan Women's University, The University of Osaka, Osaka Medical and Pharmaceutical University

Language: English

Topic: Toxin Mechanisms and Immunotoxins

Subfield: Immunology

Field: Immunology and Microbiology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $3,258

Funders: Ministry of Education, Culture, Sports, Science and Technology, Japan Society for the Promotion of Science