Clavin, a type-1 ribosome-inactivating protein from Aspergillus clavatus IFO 8605 - cDNA isolation, heterologous expression, biochemical and biological characterization of the recombinant protein

Dino Parente, Giuseppe Raucci, Bruna Celano, Aurelio Pacilli, Lorenzo Zanoni, Silvana Canevari, Elena Adobati, Maria I. Colnaghi, Franco Dosio, Silvia Arpicco, Luigi Cattel, Antonio Mele, Rita De Santis

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Abstract

We describe the cloning and expression of a new cDNA from the filamentous fungus Aspergillus clavatus IFO 8605. This cDNA contains an open reading frame (ORF) that predicts a putative ribonuclease precursor with high similarity to the α-sarcin family of ribosome-inactivating proteins (RIPs). The cDNA encoding the mature protein was expressed in Escherichia coli, and the recombinant protein, a 17-kDa polypeptide designated clavin was purified and characterized. Clavin shows typical type-1 RIP properties: specific cleavage of ribosomal and synthetic RNA and inhibition of protein synthesis in cell-free and cellular systems. When selectively targeted to a tumour cell antigen by coupling to a monoclonal antibody (mAb) clavin was able to inhibit protein synthesis at nanomolar concentration. Pharmacokinetics analysis in mice indicated an elimination half-life (t( 1/4 β)) of 7.4 h with no particular accumulation in major organs. Liver toxicity was very limited and transient while no alteration of kidney function was observed. Clavin induced a late and very low antibody response in mice. The in vitro and in vivo biological characteristics of clavin, together with its availability in large amounts, suggest the usefulness of this toxin in the production of toxic chemical conjugates.

Original languageEnglish
Pages (from-to)272-280
Number of pages9
JournalEuropean Journal of Biochemistry
Volume239
Issue number2
Publication statusPublished - 1996

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Keywords

  • Protein synthesis
  • Ribonuclease
  • Ribosome-activating protein

ASJC Scopus subject areas

  • Biochemistry

Cite this

Parente, D., Raucci, G., Celano, B., Pacilli, A., Zanoni, L., Canevari, S., Adobati, E., Colnaghi, M. I., Dosio, F., Arpicco, S., Cattel, L., Mele, A., & De Santis, R. (1996). Clavin, a type-1 ribosome-inactivating protein from Aspergillus clavatus IFO 8605 - cDNA isolation, heterologous expression, biochemical and biological characterization of the recombinant protein. European Journal of Biochemistry, 239(2), 272-280.