Mosayeb Rostamian1, Seyed Jafar Mousavy1, Firouz Ebrahimi1, Seyyed Abolghasem Ghadami2,3, Nader Sheibani4, Mohammad Ebrahim Minaei1, Mohammad Ali Arefpour Torabi1. 1. Dept. of Biology, Faculty of Basic Sciences, Imam Hussein University, Tehran, Iran. jmosavi@ihu.ac.ir 2. Dept. of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran. 3. Medical Biology Research Center, Kermanshah University of Medical Science, Kermanshah, Iran. 4. Dept. of Ophthalmology and Visual Sciences and Pharmacology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Abstract
BACKGROUND: Recently, botulinum neurotoxin (BoNT)-derived recombinant proteins have been suggested as potential botulism vaccines. Here, with concentrating on BoNT type E (BoNT/E), we studied two of these binding domain-based recombinant proteins: a multivalent chimer protein, which is composed of BoNT serotypes A, B and E binding subdomains, and a monovalent recombinant protein, which contains 93 amino acid residues from recombinant C-terminal heavy chain of BoNT/E (rBoNT/E-HCC). Both proteins have an identical region (48 aa) that contains one of the most important BoNT/E epitopes (YLTHMRD sequence). METHODS: The recombinant protein efficiency in antibody production, their structural differences, and their BoNT/E-epitope location were compared by using ELISA, circular dichroism, computational modeling, and hydrophobicity predictions. RESULTS: Immunological studies indicated that the antibody yield against rBoNT/E-HCC was higher than chimer protein. Cross ELISA confirmed that the antibodies against the chimer protein recognized rBoNT/E-HCC more efficiently. However, both antibody groups (anti-chimer and anti-rBoNT/E-HCC antibodies) were able to recognize other proteins. Structural studies with circular dichroism showed that chimer proteins have slightly more secondary structures than rBoNT/E-HCC. CONCLUSION: The immunological results suggested that the above-mentioned identical region in rBoNT/E-HCC is more exposed. Circular dichroism, computational protein modeling and hydrophobicity predictions indicated a more exposed location for the identical region in rBoNT/E-HCC than the chimer protein, which is strongly in agreement with immunological results.
BACKGROUND: Recently, botulinum neurotoxin (BoNT)-derived recombinant proteins have been suggested as potential botulism vaccines. Here, with concentrating on BoNT type E (BoNT/E), we studied two of these binding domain-based recombinant proteins: a multivalent chimer protein, which is composed of BoNT serotypes A, B and E binding subdomains, and a monovalent recombinant protein, which contains 93 amino acid residues from recombinant C-terminal heavy chain of BoNT/E (rBoNT/E-HCC). Both proteins have an identical region (48 aa) that contains one of the most important BoNT/E epitopes (YLTHMRD sequence). METHODS: The recombinant protein efficiency in antibody production, their structural differences, and their BoNT/E-epitope location were compared by using ELISA, circular dichroism, computational modeling, and hydrophobicity predictions. RESULTS: Immunological studies indicated that the antibody yield against rBoNT/E-HCC was higher than chimer protein. Cross ELISA confirmed that the antibodies against the chimer protein recognized rBoNT/E-HCC more efficiently. However, both antibody groups (anti-chimer and anti-rBoNT/E-HCC antibodies) were able to recognize other proteins. Structural studies with circular dichroism showed that chimer proteins have slightly more secondary structures than rBoNT/E-HCC. CONCLUSION: The immunological results suggested that the above-mentioned identical region in rBoNT/E-HCC is more exposed. Circular dichroism, computational protein modeling and hydrophobicity predictions indicated a more exposed location for the identical region in rBoNT/E-HCC than the chimer protein, which is strongly in agreement with immunological results.
Authors: Dylan Chivian; David E Kim; Lars Malmström; Philip Bradley; Timothy Robertson; Paul Murphy; Charles E M Strauss; Richard Bonneau; Carol A Rohl; David Baker Journal: Proteins Date: 2003
Authors: Eric F Pettersen; Thomas D Goddard; Conrad C Huang; Gregory S Couch; Daniel M Greenblatt; Elaine C Meng; Thomas E Ferrin Journal: J Comput Chem Date: 2004-10 Impact factor: 3.376
Authors: S N Beljelarskaya; O V Orlova; V L Drutsa; V A Orlov; A V Timohova; N N Koroleva; V I Popenko; A V Ivanov; P V Spirin; V S Prassolov; P M Rubtsov; S N Kochetkov Journal: Biochem Biophys Rep Date: 2016-05-30