Purification and properties of new ribosome-inactivating proteins with RNA N-glycosidase activity

Bolognesi, Andrea; Barbieri, Luigi; Abbondanza, Ada; Falasca, Anna Ida; Carnicelli, Domenica; Battelli, Maria Giulia; Stirpe, Fiorenzo

doi:10.1016/0167-4781(90)90002-j

Cited by 71 publications

(32 citation statements)

References 38 publications

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“…Pokeweed antiviral proteins (PAP's) (Irvin, 1975;lrvin et al, 1980;Barbieri et al, 1982;Bolognesi et al, 1990) are isozymes from different tissues of Phytolacca americana at different developing stages, noticed by their ability to inhibit the multiplication of virus. We now know that they are type I ribosome-inactivating proteins (RIP's) (Stirpe et al, 1992); which act as RNA N-glycosidase, removing an invariant adenine base from a loop region of rRNA interrupting and the process of protein synthesis.…”

Section: Introductionmentioning

confidence: 99%

“…Several isozymes of PAP's have been isolated including PAP from spring leaves (Irvin, 1975), PAP II from summer leaves (Irvin et aL, 1980), PAP-S from seeds (Barbieri et al, 1982), PAP-R from roots (Bolognesi et al, 1990), and PAP-C from cell culture (Barbieri et al, 1989). Although PAP's have high sequence homology (70-80%) (Kung et al, 1990;Lin et al, 1991;Kataoka et aL, 1992) and, as representatives of RIP's, are supposed to have a common catalytic mechanism, on cells or on intact ribosomes from animals, plants or bacteria, they have very different toxicities or inhibition activities.…”

Section: Introductionmentioning

confidence: 99%

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Crystallization and preliminary crystallographic analyses of pokeweed antiviral protein from seeds

Zeng

Huang

et al. 1998

Acta Crystallogr D Biol Cryst

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Pokeweed antiviral protein from seeds (PAP-S) is a ribosome inactivating protein which has lowest toxicity and highest inhibition activity as opposed to other pokeweed antiviral proteins and its three potential glycosylation sites (10, 44, 255) were shown to bind to N-acetylglucosamine. Good quality crystals of PAP-S were grown at high protein concentration (100 mg ml-~) and high temperature (306 K). The crystals have space group 1222 and cell parameters a --78.7, b = 85.2 and c --93.0 A. An X-ray diffraction data set with resolution up to 1.8 A was collected. This high-resolution data will help to locate the sugars bound to the protein and provide accurate structural data for understanding structure-function relationships of PAP-S.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Crystallization and preliminary crystallographic analyses of pokeweed antiviral protein from seeds

Zeng

Huang

et al. 1998

Acta Crystallogr D Biol Cryst

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“…Pyramidatine and ocymoidine were purified from the seeds of Saponaria ocymoides and Vaccaria pyramidata, respectively, by a modification of a previously described method (Bolognesi et al, 1990), including sequential ion-exchange chromatography on SSepharose and CM-Sepharose columns (Pharmacia, Sweden), followed by hydrophobic interaction chromatography on a phenylSepharose column (Pharmacia) for Saponaria ocymoides protein, or by filtration on Amicon PM1O membrane for Vaccaria pyramidata protein (Bolognesi et al, 1995).…”

Section: Materials and Methods Preparation Of Immunoconjugatesmentioning

confidence: 99%

Immunoconjugates made of an anti-EGF receptor monoclonal antibody and type 1 ribosome-inactivating proteins from Saponaria ocymoides or Vaccaria pyramidata

Massimo

Loreto²,

Pacilli³

et al. 1997

Br J Cancer

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Summary The present paper describes two immunoconjugates consisting of an anti-epidermal growth factor receptor (EGFR) monoclonal antibody (MAb), named Mint5, covalently linked to the type 1 ribosome-inactivating proteins (RlPs) ocymoidine (Ocy) and pyramidatine (Pyra) from Saponaria ocymoides and Vaccaria pyramidata respectively. Both antibody and toxins are shown to retain their respective biological properties upon chemical conjugation. The immunoconjugates exert specific inhibition of EGFR expressing target cell proliferation and protein synthesis in in vitro assays and also inhibit the growth of grafted human tumour cells in nude mice.Keywords: immunoconjugate; anti-epidermal growth factor receptor; ocymoidine; pyramidatineThe clinical use of immunotoxins for the treatment of cancer is currently under evaluation worldwide. The therapeutic potentiality of immunotoxins and preclinical and clinical results over the last 20 years have been reviewed recently (Thrush et al, 1996), indicating that, although immunotoxins seem promising for systemic therapy of haematological malignancies, a number of different problems still need to be solved, especially for the treatment of solid tumours. In particular, the refining of dose regimen and administration route, the combination with chemotherapy and the reduction of immunogenicity are the major goals of future research. In this paper, we describe the preparation of two new immunoconjugates made of an anti-epidermal growth factor receptor monoclonal antibody, named MintS, chemically linked to either ocymoidine or pyramidatine, toxins from Saponaria ocymoides and Vaccaria pyramidata respectively.The epidermal growth factor (EGF) and its receptor play a critical role in the growth and regulation of many normal and malignant cell types. EGFR overexpression is a common feature in most carcinomas and correlates with poor prognosis (Fox et al, 1994).The potential value of EGFR as a target for the diagnosis and therapy of human tumours has been recognized for several years, and the use of anti-EGFR monoclonal antibodies may provide therapeutic tools in the treatment of tumours overexpressing the receptor (Ennis et al, 1991).Moreover, immunoconjugates of EGFR-specific monoclonal antibodies to either gelonin (Ozawa et al, 1989) or ricin A chain (Masui et al, 1989) were shown to reduce the growth of human tumour cells transplanted into athymic mice.

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“…Pokeweed antiviral protein (PAP) is an N-glycosidase isolated from the pokeweed plant (Phytolacca americana) (Endo et al 1988;Bolognesi et al 1990). PAP is a member of a larger group of proteins referred to as ribosomeinactivating proteins, and all are characterized by their ability to depurinate the highly conserved sarcin/ricin loop of the large ribosomal RNA.…”

Section: Introductionmentioning

confidence: 99%

Pokeweed antiviral protein depurinates the sarcin/ricin loop of the rRNA prior to binding of aminoacyl-tRNA to the ribosomal A-site

Mansouri¹,

Nourollahzadeh²,

Hudak³

2006

RNA

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Ribosome-inactivating proteins, such as the pokeweed antiviral protein (PAP), inhibit translation by depurinating the conserved sarcin/ricin loop of the large ribosomal RNA. Depurinated ribosomes are unable to bind elongation factor 2, and, thus, the translocation step of the elongation cycle is inhibited. Though the consequences of depurination are well characterized, the ribosome conformation required for depurination to take place has not been described. In this report, we correlate biochemical and genetic data to conclude that pokeweed antiviral protein depurinates the sarcin/ricin loop when the A-site of the ribosomal peptidyl-transferase center is unoccupied. We show that prior incubation of ribosomes with puromycin, an analog of the 39-terminus of aminoacyl-tRNA, inhibits both binding and depurination by PAP in a concentration-dependent manner. Expression of PAP in the yeast strain mak8-1 results in little depurination unless the cells are lysed, a process that would promote loss of aminoacyl-tRNA from the ribosome. The mak8-1 strain is known to exhibit a higher affinity for aminoacyl-tRNA compared with wild-type cells, and therefore, its ribosomes are more resistant to PAP in vivo. These data contribute to the mechanism of action of pokeweed antiviral protein; specifically, they have uncovered the ribosomal conformation required for depurination that leads to subsequent translation inhibition.

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Purification and properties of new ribosome-inactivating proteins with RNA N-glycosidase activity

Cited by 71 publications

References 38 publications

Crystallization and preliminary crystallographic analyses of pokeweed antiviral protein from seeds

Crystallization and preliminary crystallographic analyses of pokeweed antiviral protein from seeds

Immunoconjugates made of an anti-EGF receptor monoclonal antibody and type 1 ribosome-inactivating proteins from Saponaria ocymoides or Vaccaria pyramidata

Pokeweed antiviral protein depurinates the sarcin/ricin loop of the rRNA prior to binding of aminoacyl-tRNA to the ribosomal A-site

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