Isolation and enzymatic characterization of lamjapin, the first ribosome-inactivating protein from cryptogamic algal plant (Laminaria japonica A).
Lamjapin, a novel type Iota ribosome-inactivating protein, has been isolated from kelp (Laminaria japonica A), a marine alga. This protein has been extensively purified through multiple chromatography columns. With a molecular mass of approximately 36 kDa, lamjapin is slightly larger than the other known single-chain ribosome-inactivating proteins from the higher plants. Lamjapin can inhibit protein synthesis in rabbit reticulocyte lysate with an IC50 of 0.69 nm. It can depurinate at multiple sites of RNA in rat ribosome and produce the diagnostic R-fragment and three additional larger fragments after the aniline reaction. Lamjapin can deadenylate specifically at the site A20 of the synthetic oligoribonucleotide (35-mer) substrate that mimics the sarcin/ricin domain (SRD) of rat ribosomal 28S RNA. However, it cannot hydrolyze the N-C glycosidic bond of guanosine, cytidine or uridine at the corresponding site of the A20 of three mutant SRD RNAs. Lamjapin exhibits the same base and position requirement as the ribosome-inactivating proteins from higher plants. We conclude that lamjapin is an RNA N-glycosidase that belongs to the ribosome-inactivating protein family. This study reports for the first time that ribosome-inactivating protein exists in the lower cryptogamic algal plant.
D000818:Animals; D001483:Base Sequence; D002474:Cell-Free System; D066298:In Vitro Techniques; D007795:Laminaria / Q000737:chemistry*; D008970:Molecular Weight; D009699:N-Glycosyl Hydrolases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D012335:RNA, Ribosomal / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism; D054788:Ribosome Inactivating Proteins; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity
D010940:Plant Proteins; D011500:Protein Synthesis Inhibitors; D012335:RNA, Ribosomal; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins