TY - JOUR
T1 - Implications of high-molecular-weight oligomers of the binary toxin from Bacillus sphaericus
AU - Smith, A. W.
AU - Cámara-Artigas, A.
AU - Brune, D. C.
AU - Allen, James
N1 - Funding Information:
The authors would like to thank Elizabeth Davidson for providing the B. sphaericus strains, Jeff LaBelle for help with the dynamic light scattering experiments, and JoAnn Williams for careful reading of the manuscript. A.C.A. is supported by Grant BIO-2003 04274 from the Dirección General Científica y Técnica, Ministerio de Ciencia y Tecnología, Spain. The mass spectrometer was purchased with funds provided by NSF CHE-0131222.
PY - 2005/1
Y1 - 2005/1
N2 - The mosquito-larvicidal binary toxin produced by Bacillus sphaericus is composed of BinB and BinA, which have calculated molecular weights of 51.4 and 41.9 kDa, respectively. NaOH extracts of B. sphaericus spores were analyzed using SDS-PAGE. Stained gels showed bands with molecular weights corresponding to those of BinB and BinA as well as two additional bands at 110 and 125 kDa. The matrix-assisted laser desorption/ionization mass spectrum of the purified 110 and 125 kDa bands showed two peaks at 104,160 and 87,358 Da that are assigned to dimers of BinB and BinA, respectively. Mass spectral analysis of trypsin-digested 110 and 125 kDa bands showed peaks at 51,328, 43,523, 43,130, and 40,832 Da that assigned to undigested BinB, two forms of digested BinB and digested BinA, respectively. Dynamic light scattering studies showed a solution of the purified 110 and 125 kDa bands was comprised almost entirely (99.6% of total mass) of a particle with a hydrodynamic radius of 5.6 ± 1.2 nm and a calculated molecular weight of 186 ± 38 kDa. These data demonstrate that the binary toxin extracted from B. sphaericus spores can exist in solution as an oligomer containing two copies each of BinB and BinA.
AB - The mosquito-larvicidal binary toxin produced by Bacillus sphaericus is composed of BinB and BinA, which have calculated molecular weights of 51.4 and 41.9 kDa, respectively. NaOH extracts of B. sphaericus spores were analyzed using SDS-PAGE. Stained gels showed bands with molecular weights corresponding to those of BinB and BinA as well as two additional bands at 110 and 125 kDa. The matrix-assisted laser desorption/ionization mass spectrum of the purified 110 and 125 kDa bands showed two peaks at 104,160 and 87,358 Da that are assigned to dimers of BinB and BinA, respectively. Mass spectral analysis of trypsin-digested 110 and 125 kDa bands showed peaks at 51,328, 43,523, 43,130, and 40,832 Da that assigned to undigested BinB, two forms of digested BinB and digested BinA, respectively. Dynamic light scattering studies showed a solution of the purified 110 and 125 kDa bands was comprised almost entirely (99.6% of total mass) of a particle with a hydrodynamic radius of 5.6 ± 1.2 nm and a calculated molecular weight of 186 ± 38 kDa. These data demonstrate that the binary toxin extracted from B. sphaericus spores can exist in solution as an oligomer containing two copies each of BinB and BinA.
KW - AB toxin
KW - Anopheles
KW - Culex
KW - Dynamic light scattering
KW - Larvicide
KW - Mass spectrometry
KW - Mosquitocidal
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U2 - 10.1016/j.jip.2004.10.005
DO - 10.1016/j.jip.2004.10.005
M3 - Article
C2 - 15707866
AN - SCOPUS:13444262448
SN - 0022-2011
VL - 88
SP - 27
EP - 33
JO - Journal of Invertebrate Pathology
JF - Journal of Invertebrate Pathology
IS - 1
ER -