title: Maize ribosome-inactivating protein uses Lys158-lys161 to interact with ribosomal protein P2 and the strength of interaction is correlated to the biological activities.
abstract: Ribosome-inactivating proteins (RIPs) inactivate prokaryotic or eukaryotic ribosomes by removing a single adenine in the large ribosomal RNA. Here we show maize RIP (MOD), an atypical RIP with an internal inactivation loop, interacts with the ribosomal stalk protein P2 via Lys158-Lys161, which is located in the N-terminal domain and at the base of its internal loop. Due to subtle differences in the structure of maize RIP, hydrophobic interaction with the 'FGLFD' motif of P2 is not as evidenced in MOD-P2 interaction. As a result, interaction of P2 with MOD was weaker than those with trichosanthin and shiga toxin A as reflected by the dissociation constants (K(D)) of their interaction, which are 1037.50 ± 65.75 µM, 611.70 ± 28.13 µM and 194.84 ± 9.47 µM respectively.Despite MOD and TCS target at the same ribosomal protein P2, MOD was found 48 and 10 folds less potent than trichosanthin in ribosome depurination and cytotoxicity to 293T cells respectively, implicating the strength of interaction between RIPs and ribosomal proteins is important for the biological activity of RIPs. Our work illustrates the flexibility on the docking of RIPs on ribosomal proteins for targeting the sarcin-ricin loop and the importance of protein-protein interaction for ribosome-inactivating activity.
authors.affiliation: Biochemistry Programme and Centre for Protein Science and Crystallography, School of Life Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
authors.forename: Yuen-Ting
authors.initials: YT
authors.lastname: Wong
authors.forename: Yiu-Ming
authors.initials: YM
authors.lastname: Ng
authors.forename: Amanda Nga-Sze
authors.initials: AN
authors.lastname: Mak
authors.forename: Kong-Hung
authors.initials: KH
authors.lastname: Sze
authors.forename: Kam-Bo
authors.initials: KB
authors.lastname: Wong
authors.forename: Pang-Chui
authors.initials: PC
authors.lastname: Shaw
mesh_terms: D000818:Animals; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism; D051381:Rats; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D054788:Ribosome Inactivating Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*; D022621:Shiga Toxin / Q000235:genetics / Q000378:metabolism; D015978:Trichosanthin / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000235:genetics / Q000378:metabolism*
chemical_list: D010750:Phosphoproteins; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C493898:Shiga toxin subunit A; C031937:phosphoprotein P2, ribosomal; D015978:Trichosanthin; D022621:Shiga Toxin; D054788:Ribosome Inactivating Proteins