PMID: 9443374Aug 1, 1997Paper

The heliothis virescens 170 kDa aminopeptidase functions as "receptor A" by mediating specific Bacillus thuringiensis Cry1A delta-endotoxin binding and pore formation

Insect Biochemistry and Molecular Biology
K LuoMichael J Adang

Abstract

The relationship between Bacillus thuringiensis Cry1Aa, Cry1Ab and Cry1Ac delta-endotoxin binding and pore formation was investigated using a purified 170 kDa aminopeptidase N (APN) from Heliothis virescens brush border membranes. Aminopeptidases with molecular sizes of 110, 140 and 170 kDa were eluted from a Cry1Ac toxin affinity column using N-acetylgalactosamine. The 140 kDa aminopeptidase has a cross-reacting determinant typical of a cleaved glycosyl-phosphatidylinositol anchor. After mild base treatment to de-acylate the glycosyl-phosphatidylinositol linkage and incubation in phosphatidyl inositol phospholipase C, anti-cross-reacting determinant antibody recognized the 170 kDa protein. Kinetic binding characteristics of Cry1A toxins to purified 170 kDa APN were determined using surface plasmon resonance. Cry1Aa, Cry1Ab and Cry1Ac, but not Cry1C and Cry1E toxins recognized 170 kDa APN. Each Cry1A toxin recognized two binding sites: a high affinity site with KD ranging from 41 to 95 nM and a lower affinity site with KD in the 325 to 623 nM range. N-acetylgalactosamine inhibited Cry1Ac but not Cry1Aa and Cry1Ab binding to 170 kDa APN. When reconstituted into phospholipid vesicles, the 170 kDa APN promoted toxin-induced 86Rb+ ...Continue Reading

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Citations

Aug 27, 2011·Applied Microbiology and Biotechnology·Santosh Kumar UpadhyayPradhyumna Kumar Singh
May 1, 2007·Protein Expression and Purification·Ravinder KaurRaj Bhatnagar
Dec 13, 2005·Biochemical and Biophysical Research Communications·Mi Kyong LeeJeng-Shong Chen
Jun 27, 2006·Journal of Invertebrate Pathology·Juan Luis Jurat-Fuentes, Michael J Adang
Jun 27, 2006·Journal of Invertebrate Pathology·Karlygash G AimanovaSarjeet S Gill
Nov 25, 2003·Archives of Insect Biochemistry and Physiology·Mark E Whalon, Byron A Wingerd
Oct 3, 2001·Nature Biotechnology·B E Tabashnik
Jul 22, 2004·European Journal of Biochemistry·Juan L Jurat-Fuentes, Michael J Adang
May 1, 2004·Critical Reviews in Microbiology·Abanti ChattopadhyayRakesh Bhatnagar
Oct 5, 2007·Molecular Biotechnology·Hiroshi IshikawaRyoichi Sato
Jun 11, 2011·Biotechnology Letters·Santosh K Upadhyay, Pradhyumna K Singh
May 25, 2015·Parasites & Vectors·Emmanuel Dunstand-GuzmánFernando Iván Flores-Pérez
Mar 28, 2002·Applied and Environmental Microbiology·Androulla GillilandDavid J Ellar
Jun 8, 2007·Microbiology and Molecular Biology Reviews : MMBR·Craig R Pigott, David J Ellar
Jan 3, 2001·Applied and Environmental Microbiology·J L Jurat-Fuentes, M J Adang
Jul 8, 2005·Applied and Environmental Microbiology·Shogo AtsumiRyoichi Sato

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