scholarly journals Gene cloning and construction of prokaryotic and plant expression vectors of RICIN-A-Chain/PAP-S1 fusion protein and its inhibition of protein synthesis

2016 ◽  
Author(s):  
Yasser S. Hassan ◽  
Sherry L. Ogg
Keyword(s):  
Fusion Protein ◽  
Expression Vectors ◽  
Single Chain ◽  
Antiviral Protein ◽  
Ricin A Chain ◽  
C Terminus ◽  
N Terminus ◽  
A Cell ◽  
A Chain ◽  
Ricin A

AbstractPokeweed antiviral protein (PAP) is a single-chain ribosome-inactivating protein that exists in several forms isolated from various organs and at different stages of development of Phytolacca americana (pokeweed). In this study, PAP-S1, one of the two known isoforms found in seeds, was isolated and PCR amplified using primers based on the known mRNA of PAP-S2, the other known form found in seeds. The complete cDNA encoding PAP-S1 was determined here for the first time. PAP-S1 is a potent antiviral protein with many potential clinical applications. However, it was found to be dosage dependent with observed side effects at high dosage. In this study, we report the production of a recombinant antiviral peptide-fusion protein between Ricin A-chain and PAP-S1. The peptide-fusion recombinant proteins Ricin-A-Chain/PAP-S1 and PAP-S1/Ricin-A-Chain were generated by joining the Nterminus of PAP-S1 to the C-terminus of Ricin A-chain and the C-terminus of PAP-S1 to the N-terminus of Ricin A-chain respectively, and were expressed in an Escherichia coli cell free expression systems. The peptide-fusion recombinant protein Ricin-A-Chain/PAP-S1 (F2) was found to be more active than the PAPS1/Ricin-A-chain (F1) and similar to PAP-S1 in a cell free prokaryotic environment, and both showed much stronger activity in a cell free eukaryotic environment. The DNA sequence of the complete cDNA of PAP-S1 and of the peptide-fusion protein Ricin-A-Chain/PAP-S1 with the PAP-S1 signal peptide at the N-terminus of Ricin Achain were inserted in plant destination binary vectors for A. tumefaciens mediated transformation. It is the authors’ opinion that additional research should be done in order to determine both cytotoxicity and selectivity of fusion protein F2 compared to PAP-S1, as it could be a viable, more potent and less cytotoxic alternative to PAPS1 alone at high dosage, for both agricultural and therapeutic applications.

scholarly journals Novel Binding Mechanisms of Fusion Broad Range Anti-Infective Protein Ricin A Chain Mutant-Pokeweed Antiviral Protein 1 (RTAM-PAP1) against SARS-CoV-2 Key Proteins in Silico

Toxins ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 602
Author(s):  
Yasser Hassan ◽  
Sherry Ogg ◽  
Hui Ge
Keyword(s):  
In Silico ◽  
Protein Structures ◽  
Ligand Docking ◽  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Ricin A Chain ◽  
In Silico Docking ◽  
A Chain ◽  
Binding Mechanisms ◽  
Ricin A

The deadly pandemic named COVID-19, caused by a new coronavirus (SARS-CoV-2), emerged in 2019 and is still spreading globally at a dangerous pace. As of today, there are no proven vaccines, therapies, or even strategies to fight off this virus. Here, we describe the in silico docking results of a novel broad range anti-infective fusion protein RTAM-PAP1 against the various key proteins of SARS-CoV-2 using the latest protein-ligand docking software. RTAM-PAP1 was compared against the SARS-CoV-2 B38 antibody, ricin A chain, a pokeweed antiviral protein from leaves, and the lectin griffithsin using the special CoDockPP COVID-19 version. These experiments revealed novel binding mechanisms of RTAM-PAP1 with a high affinity to numerous SARS-CoV-2 key proteins. RTAM-PAP1 was further characterized in a preliminary toxicity study in mice and was found to be a potential therapeutic candidate. These findings might lead to the discovery of novel SARS-CoV-2 targets and therapeutic protein structures with outstanding functions.

scholarly journals Cytotoxicity of a cell-reactive immunotoxin containing ricin A chain is potentiated by an anti-immunotoxin containing ricin B chain.

1984 ◽  
Vol 160 (1) ◽  
pp. 341-346 ◽  
Author(s):  
E S Vitetta ◽  
R J Fulton ◽  
J W Uhr
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Ricin A Chain ◽  
Daudi Cell ◽  
A Cell ◽  
A Chain ◽  
Ricin A

In vitro killing of the human Daudi cell line by either univalent [F(ab')] or divalent (IgG) forms of rabbit anti-human Ig (RAHIg) coupled to ricin A chain can be specifically potentiated by a "piggyback" treatment with ricin B chain coupled to goat anti-rabbit Ig (GARIg). When cells are treated with univalent immunotoxin (IT) [F(ab') RAHIg-A] and then cultured, IT can be detected on the cell surface for at least 5 h, since GARIg-B can still enhance killing at this time. These results provide a strategy for in vivo use of A chain- and B chain-containing IT.

scholarly journals Major Structural Differences between Pokeweed Antiviral Protein and Ricin A-Chain do not Account for their Differing Ribosome Specificity

1996 ◽  
Vol 235 (1-2) ◽  
pp. 159-166 ◽  
Author(s):  
John A. Chaddock ◽  
Arthur F. Monzingo ◽  
Jon D. Robertus ◽  
J. Michael Lord ◽  
Lynne M. Roberts
Keyword(s):  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Ricin A Chain ◽  
Structural Differences ◽  
A Chain ◽  
Ricin A

scholarly journals Cytotoxicity of a recombinant ricin-A-chain fusion protein containing a proteolytically-cleavable spacer sequence

FEBS Letters ◽  
1990 ◽  
Vol 273 (1-2) ◽  
pp. 200-204 ◽  
Author(s):  
Mary O'Hare ◽  
Alex N. Brown ◽  
Khalid Hussain ◽  
Angelika Gebhardt ◽  
Graham Watson ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Ricin A Chain ◽  
A Chain ◽  
Ricin A

scholarly journals Novel anti-SARS-CoV-2 mechanisms of fusion broad range anti- infective protein ricin A chain mutant-pokeweed antiviral protein 1 (RTAM-PAP1) in silico

2020 ◽  
Author(s):  
Yasser Hassan ◽  
Sherry Ogg ◽  
Hui Ge
Keyword(s):  
In Silico ◽  
Protein Structures ◽  
Ligand Docking ◽  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Ricin A Chain ◽  
High Affinity ◽  
A Chain ◽  
Binding Mechanisms ◽  
Ricin A

Abstract A deadly pandemic named COVID-19 caused by a new coronavirus SARS- CoV-2 has emerged in 2019 and is still spreading globally at a dangerous pace. As of today, there are no proven vaccines, therapies or even strategies to fight off this virus. Here, we describe the in silico results of a novel broad range anti-infective fusion protein RTAM-PAP1 against the various key pro- teins of SARS-CoV-2 using the latest protein-ligand docking software. RTAM-PAP1 was compared against the SARS-CoV-2 B38 antibody, ricin A chain, pokeweed antiviral protein from leaves and the lectin griffithsin using CoDockPP special COVID-19 version. These experiments revealed novel binding mechanisms of RTAM-PAP1 with high affinity to numerous targets with anti-SARS-CoV-2 effects. RTAM-PAP1 was further characterized in a preliminary toxicity study in mice and was found to likely be a potent anti- SARS-CoV-2 agent. These findings might lead to the discovery of novel SARS-CoV-2 targets and therapeutic protein structures.

Inductive regulation of cell fusion in leech

Development ◽  
1999 ◽  
Vol 126 (15) ◽  
pp. 3381-3390 ◽  
Author(s):  
D.E. Isaksen ◽  
N.J. Liu ◽  
D.A. Weisblat
Keyword(s):  
Cell Fusion ◽  
Critical Period ◽  
Ribonuclease A ◽  
Fusion Event ◽  
Ricin A Chain ◽  
Helobdella Robusta ◽  
A Cell ◽  
A Chain ◽  
Ricin A ◽  
Cell Cell

Cell-cell fusion is a component of many different developmental processes, but little is known about how cell-cell fusion is regulated. Here we investigate the regulation of a stereotyped cell-cell fusion event that occurs among the endodermal precursor cells of the glossiphoniid leech Helobdella robusta. We find that this fusion event is regulated inductively by a cell that does not itself fuse. We also show that biochemical arrest (by microinjection with ricin A chain or ribonuclease A) of the inducer or either of the fusion partners prevents fusion, but only if the arrest is initiated during a critical period long before the time at which fusion normally occurs. If the arrest occurs after this critical period, fusion occurs on schedule. These results suggest that both fusion partners play active roles in the process and that neither the induction nor the fusion itself requires concomitant protein synthesis.

Selective killing of smooth muscle cells in culture by the ricin A-chain conjugated with monoclonal antibodies to a cell surface antigen via a dextran bridge

1985 ◽  
Vol 41 (10) ◽  
pp. 1342-1344 ◽  
Author(s):  
O. Yu. Printseva ◽  
A. I. Faerman ◽  
A. V. Maksimenko ◽  
A. G. Tonevitsky ◽  
O. B. Ilynsky ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Monoclonal Antibodies ◽  
Cell Surface ◽  
Smooth Muscle Cells ◽  
Surface Antigen ◽  
Muscle Cells ◽  
Ricin A Chain ◽  
A Cell ◽  
A Chain ◽  
Ricin A

scholarly journals Novel anti-SARS-CoV-2 mechanisms of fusion broad range anti- infective protein ricin A chain mutant-pokeweed antiviral protein 1 (RTAM-PAP1) in silico

2020 ◽  
Author(s):  
Yasser Hassan ◽  
Sherry Ogg ◽  
Hui Ge
Keyword(s):  
In Silico ◽  
Protein Structures ◽  
Ligand Docking ◽  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Ricin A Chain ◽  
High Affinity ◽  
A Chain ◽  
Binding Mechanisms ◽  
Ricin A

Abstract A deadly pandemic named COVID-19 caused by a new coronavirus SARS- CoV-2 has emerged in 2019 and is still spreading globally at a dangerous pace. As of today, there are no proven vaccines, therapies or even strategies to fight off this virus. Here, we describe the in silico results of a novel broad range anti-infective fusion protein RTAM-PAP1 against the various key pro- teins of SARS-CoV-2 using the latest protein-ligand docking software. RTAM-PAP1 was compared against the SARS-CoV-2 B38 antibody, ricin A chain, pokeweed antiviral protein from leaves and the lectin griffithsin using CoDockPP special COVID-19 version. These experiments revealed novel binding mechanisms of RTAM-PAP1 with high affinity to numerous targets with anti-SARS-CoV-2 effects. RTAM-PAP1 was further characterized in a preliminary toxicity study in mice and was found to likely be a potent anti- SARS-CoV-2 agent. These findings might lead to the discovery of novel SARS-CoV-2 targets and therapeutic protein structures.

scholarly journals Human ribosomal P1-P2 heterodimer represents an optimal docking site for ricin A chain with a prominent role for P1 C-terminus

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Przemysław Grela ◽  
Xiao-Ping Li ◽  
Patrycja Horbowicz ◽  
Monika Dźwierzyńska ◽  
Marek Tchórzewski ◽  
...  
Keyword(s):  
Docking Site ◽  
Ricin A Chain ◽  
C Terminus ◽  
A Chain ◽  
Ricin A

Shiga toxin 1 and ricin A chain bind to human polymorphonuclear leucocytes through a common receptor

2010 ◽  
Vol 432 (1) ◽  
pp. 173-180 ◽  
Author(s):  
Valentina Arfilli ◽  
Domenica Carnicelli ◽  
Laura Rocchi ◽  
Francesca Ricci ◽  
Pasqualepaolo Pagliaro ◽  
...  
Keyword(s):  
Binding Sites ◽  
Biological Significance ◽  
Mannose Receptor ◽  
Polymorphonuclear Leucocytes ◽  
Single Chain ◽  
Ricin A Chain ◽  
Common Receptor ◽  
Intestinal Infections ◽  
A Chain ◽  
Ricin A

The main cause of acute renal failure in children is HUS (haemolytic uraemic syndrome), a consequence of intestinal infections with Escherichia coli strains producing Stx (Shiga toxins). Stx released in the gut by the non-invasive bacteria reach the bloodstream and are targeted to cerebral and renal endothelium triggering HUS. PMN (polymorphonuclear leucocytes) seem to be involved in Stx delivery through an unidentified membrane receptor (Kd=10−8 M; 2×105 binding sites) which does not allow internalization. Some experts in the field have defined the Stx–PMN interaction as non-specific and of little biological significance. In the present study, we show that the A chain of ricin, the well-known plant RIP (ribosome-inactivating protein), interacts with PMN (Kd=10−9 M; 2×105 binding sites) competing for the same receptor that recognizes Stx, whereas diphtheria toxin and several agonists of TLRs (Toll-like receptors) or the mannose receptor were ineffective. No toxic effects of ricin A chain on PMN were observed, as assessed by measuring protein synthesis and the rate of spontaneous apoptosis of leucocytes. Moreover, two single-chain RIPs (gelonin and saporin S6) had the same competing effect. Thus RIPs and Stx1 share structural similarities, the same enzymatic activity and a common receptor on PMN. These observations reveal that the Stx–PMN interaction is specific, confirming that PMN recognize molecular patterns common to different foreign molecules.

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