Ebulin-RP, a novel member of the Ebulin gene family with low cytotoxicity as a result of deficient sugar binding domains

• Published in: Biochimica et biophysica acta Vol. 1862; no. 3; pp. 460 - 473
• Main Authors: Iglesias, Rosario, Ferreras, J. Miguel, Di Maro, Antimo, Citores, Lucía
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2018
• Subjects: amino acid sequences; Apoptosis; carbohydrate binding; complementary DNA; cytotoxicity; enzyme activity; genes; leaves; Lectin; lectins; moieties; N-glycoside hydrolases; neoplasm cells; neoplasms; ribosomal RNA; Ribosome-inactivating protein (RIP); Ricin; rRNA N-glycosidase; Sambucus ebulus; Sambucus ebulus L; sugars; Lectin; rRNA N-glycosidase; Sambucus ebulus L; Ricin; Ribosome-inactivating protein (RIP); Apoptosis
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2017.11.014

Sambucus ebulus is a rich source of ribosome-inactivating proteins (RIPs) and RIP-related lectins generated from multiple genes. These proteins differ in their structure, enzymatic activity and sugar binding specificity. We have purified and characterized ebulin-RP from S. ebulus leaves and determined the amino acid sequence by cDNA cloning. Cytotoxicity was studied in a variety of cancer cells and a comparative study of the ability of ebulin-RP to bind sugars using “in vitro” and “in silico” approaches was performed. Ebulin-RP is a novel heterodimeric type 2 RIP present in S. ebulus leaves together with the type 2 RIP ebulin l, which displayed rRNA N-glycosidase activity but unlike ebulin l, lacked functional sugar binding domains. As a consequence of changes in its B-chain, ebulin-RP displayed lower cytotoxicity than ebulin l towards cancer cells and induced apoptosis as the predominant pattern of cell death. Ebulin-RP is a novel member of the ebulin gene family with low cytotoxicity as a result of deficient sugar binding domains. Type 2 RIP genes from Sambucus have evolved to render proteins with different sugar affinities that may be related to different biological activities and could result in an advantage for the plant. The ebulin family of RIPs and lectins can serve as a good model for studying the evolutionary process which may have occurred in RIPs. The lack of cytotoxicity of ebulin-RP makes it a good candidate as a toxic moiety in the construction of immunotoxins and conjugates directed against specific targets. [Display omitted] •Ebulin-RP is a new type 2 RIP from S. ebulus leaves related to ebulin l and SEA.•Ebulin-RP is enzymatically similar to type 2 RIPs such as ebulin l and ricin.•Ebulin-RP fails to agglutinate erythrocytes or to bind AT-Sepharose 6B.•Ebulin-RP displays low cytotoxicity as a result of deficient sugar binding domains.•Ebulin-RP induces apoptosis as the predominant pattern of cell death.