two disulfide mutants in domain i of bacillus thuringiensis cry3aa δ-endotoxin increase stability with no effect on toxicity两个二硫化苏云金杆菌的突变体在域我cry3aaδ-endotoxin对毒性增加稳定性没有影响.pdf

Advances in Biological Chemistry, 2012, 2, 123-131 ABC /10.4236/abc.2012.22015 Published Online May 2012 (http://www.SciRP.org/journal/abc/) Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa -endotoxin increase stability with no effect on * toxicity 1 2 1 1# 3,4,5,# Sheng-Jiun Wu , Alvaro M. Florez , Bradley J. Homoelle , Donald H. Dean , Oscar Alzate 1Biochemistry Department, Ohio State University, Columbus, USA 2Laboratorio de Biología Molecular y Biotecnología, Facultad de Medicina, Universidad de Santander (UDES), Bucaramanga, Co- lombia 3Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, USA 4Universidad Pontificia Bolivariana, Medellín, Colombia 5Biophysics Program, Ohio State University, Columbus, USA Email: #alzate@, #dean.10@ Received 28 January 2012; revised 1 March 2012; accepted 10 March 2012 ABSTRACT Keywords: Disulfide Bonds; CD Spectra; Cry3Aa; Site Directed Mutagenesis To increase protein stability and test protein function, three double-cysteine mutations were individually introduced by protein engineering into the cysteine- 1. INTRODUCTION free Cry3Aa δ-endotoxin from Bacillus thuringiensis. Protein engineering is a powerful tool for modifying the These mutations were designed to create disulfide properties of polypeptide molecules. One particular ap- bonds between α-helices 2 and 5 (positions 110 - 193), plication of pr