Marnix Peferoen - Academia.edu (original) (raw)
Papers by Marnix Peferoen
Physiological Entomology, 1982
ABSTRACT The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-... more ABSTRACT The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-h photophase) Leptinotarsa decemlineata Say adults was characterized by high performance liquid chromatography, which revealed that for short-day beetles ecdysterone and ecdysone are the major constituents of the free ecdysteroids, whereas in long-day beetles products co-migrating with 2-deoxyecdysone and 2-deoxyecdysterone are just as abundant. Ecdysteroids were detected (15 ng/ml) only in the haemolymph of females. Ligation experiments showed that the induction of the diapause syndrome by inactivation of the corpora allata or the brain not only resulted in the formation of the typical diapause proteins but also in an increase of the ecdysteroid activity. It is suggested that adult diapause in L.decemlineata is regulated by a multifactorial system.
Journal of Cellular Biochemistry, 1986
ACS Symposium Series, 1992
66th International Symposium on Crop Protection, 2014
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1986
Isolated ovaries of egg laying females synthesize and secrete three yolk proteins (two vitellogen... more Isolated ovaries of egg laying females synthesize and secrete three yolk proteins (two vitellogenins and chromoprotein 2). 2. The contribution of ovarian tissue to total yolk protein production is very small, the major site of synthesis of the three yolk proteins being the fat body. 3. There is a time lag between yolk protein synthesis by the fat body and yolk protein sequestration by the ovary. 4. In egg laying females, within 1 hr after the synthesis of both vitellogenins by the fat body, they appear in the oocytes as vitellins.
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1982
The 2 vitellogenins, VFPI and VFP2, are immunologically unrelated. 2. Both the viteUogenins are l... more The 2 vitellogenins, VFPI and VFP2, are immunologically unrelated. 2. Both the viteUogenins are lipo-and glycoproteins and consist of 4 and 6 polypeptides respectively. 3. The selective uptake of proteins (2 vitellogenins and 1 chromoprotein) into the oocyte, loads them with more negative charges, which is apparently the only change. 4. In prediapausing beetles, 3 immunologically unrelated proteins accumulate. One of these diapause proteins, DP1, also accumulates in larval and pupal hemolymph. 5. The diapause proteins are all glycoproteins, consisting of one single polypeptide.
Insect Biochemistry, 1984
The accessory reproductive glands (ARGs) of the male Colorado beetle secrete about 50 polypeptide... more The accessory reproductive glands (ARGs) of the male Colorado beetle secrete about 50 polypeptides with molecular weights varying between 15,000 and 650,000. Most of these polypeptides are synthesized by the gland itself. However, some secretory polypeptides originate from the fat body and are transported to the ARGs by the haemolymph. One of these extraglandular secretory polypeptides corresponds with one of the two subunits of the major haemolymph lipoprotein. The second subunit of this lipoprotein is not secreted, but is accumulated at the microvillar zone of the secretory cells.
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1986
Two vitellogenins and chromoprotein 2 are selectively accumulated by the oocyte and cannot be det... more Two vitellogenins and chromoprotein 2 are selectively accumulated by the oocyte and cannot be detected either in follicle cells or in the germarium. 2. At the start of their accumulation in terminal oocytes they are asymmetrically distributed. 3. Endocytosis of vitellogenin l starts somewhat later than the uptake of vitellogenin 2 and chromoprotein 2. 4. In follicle cells of young follicles, a protein (DLP), immunologically related to diapause protein l, is highly concentrated. 5. During vitellogenesis DLP is sequestered by the oocytes. 6. The protein rich globules in terminal oocytes contain the vitellins as well as chromoprotein 2 and DLP.
Journal of Insect Physiology, 1982
ABSTRACT
FEMS Microbiology Letters, 1989
Canadian Journal of Microbiology, 1996
Light microscopy was used to investigate the relation between toxicity, cytopathological effects,... more Light microscopy was used to investigate the relation between toxicity, cytopathological effects, and in vivo binding of Bacillus thuringiensis CrylA(b) and CrylE toxin proteins in larvae of Lymantria dispar, Choristoneura fumiferana, Actebia fennica, and Bombyx mori. These target insects were selected for their contrasting susceptibility to the two toxins. Lymantria dispar is susceptible to CrylA(b), B. mori is susceptible to CrylE, C. fumiferana is susceptible to both, and A. fennica is not susceptible to either. In the susceptible species, both toxins caused typical pathological changes in midgut epithelial cells, including disruption and shedding of the brush border membrane, vacuolization of the cytoplasm, and swelling of the cells and their nuclei, followed by disintegration and release of cytoplasmic content into the lumen. In the highly resistant A. fennica, no cell damage was observed, but the midguts of toxin-fed larvae had a shrunken appearance. Immunohistochemical staini...
Applied and Environmental Microbiology, 1993
According to the information supplied by the manufacturer of the biotinylation kit (RPN28), the c... more According to the information supplied by the manufacturer of the biotinylation kit (RPN28), the concentration of the N-hydroxysuccinimide-biotin ester was 0.46 mg/ml. However, recently we were told that this information was incorrect and that the actual concentration was 5 mg/ml. Therefore, the molar reaction ratios of biotin molecules to insecticidal crystal protein molecules in our paper should be multiplied by 10.9. (Note that these biotinylation ratios refer to the biotin-to-protein ratio during biotinylation [Anal. Biochem 195:214-219, 1991] rather than to the number of biotins actually attached.
Pest Management in Rice, 1990
During recent years several procedures have been developed to introduce foreign genes into plant ... more During recent years several procedures have been developed to introduce foreign genes into plant cells and to subsequently regenerate fertile plants. In combination with an increasing understanding of plant gene regulation and the processing and targeting of proteins this has provided the opportunity to obtain genetically modified plants. Traits of agronomic importance can be introduced into crop plants, one example being the engineering of plants resistant to insect attack through the expression of insecticidal proteins. At present the preferential source of such insecticidal proteins is the bacterium Bacillus thuringiensis (B.t.).
European Journal of Biochemistry, 1997
We report the purification, cloning and characterization of an aminopeptidase N from the midgut e... more We report the purification, cloning and characterization of an aminopeptidase N from the midgut epithelium of Manduca sexta that binds Cry1 Ab5, an insecticidal crystal protein [ICP] from Bacillus fhuringiensis. Sequence information derived from this M. sexta aminopeptidase N was used for the cloning o f an aminopeptidase N from the midgut brush-border membrane of Plutrlla xylostella, an insect species of which some populations acquired resistance against Cry1 Ab5. Affinity chromatography on a CrylAb5 matrix was used to isolate a 220-kDa glycoprotein from the larval midgut of the lepidopteran M. sexta. On ligand blots the purified 120-kDa protein discriminates between the lepidopteran-specific Cry1 Ab5 and the coleopteran-specific Cry3A d-endotoxin. Internal amino acid sequences from the 120-kDa protein were used for the design of degenerate oligonucleotides. From a nested PCR with M. sexta midgut cDNA as template, a DNA fragment was obtained which shows similarity to prokaryotic and eukaryotic aminopeptidase N genes. This PCR fragment was used to screen cDNA libraries of larval midguts from M. sexta and P. xylostella. From the M. sextu midgut cDNA library a 2973-bp nucleotide sequence was cloned. The ORF of the sequence encodes a 942-residue aminopeptidase N (M. sexta Apn2) containing two hydrophobic regions. The NH,-terminal hydrophobic region corresponds to a secretory signal sequence and the COOH-terminal hydrophobic region is typical of glycosylphosphatidylinositol (glycosyl-PtdTns)-anchored proteins. Low-stringency hybridization of the tl xylostella midgut cDNA library with M. sexta u p 2 probes enabled the isolation of a 3118-bp sequence with an ORF encoding a 946-residue preproprotein. This aminopeptidase N (P xylostella Apnl) displays 61 % amino acid identity to M. sexta Apn2 and contains a COOH-terminal signal peptide for glycosyl-PtdIns anchor addition. Both M. sexta Apn2 and P. .xy'lostella Apnl contain four Cys residues, which are highly conserved among eukaryotic aminopeptidase N molecules. Treatment of Sf9 cells expressing the P xylostellu apnl gene with PtdIns-specific phospholipase C demonstrated that tl xylostella Apnl is attached to the insect cell tnembrane by a glycosyl-Ptdlns anchor.
Proceedings of the National Academy of Sciences, 1991
The biochemical mechanism for resistance to Bacillus thuringiensis crystal proteins was studied i... more The biochemical mechanism for resistance to Bacillus thuringiensis crystal proteins was studied in a field population of diamondback moths (Plutella xylostella) with a reduced susceptibility to the bioinsecticidal spray. The toxicity and binding characteristics of three crystal proteins [CryIA(b), CryIB, and CryIC] were compared between the field population and a laboratory strain. The field population proved resistant (greater than 200-fold compared with the laboratory strain) to CryIA(b), one of the crystal proteins in the insecticidal formulation. Binding studies showed that the two strains differ in a membrane receptor that recognizes CryIA(b). This crystal protein did not bind to the brush-border membrane of the midgut epithelial cells of the field population, either because of strongly reduced binding affinity or because of the complete absence of the receptor molecule. Both strains proved fully susceptible to the CryIB and CryIC crystal proteins, which were not present in the...
Molecular Microbiology, 1991
The insecticidal crystal proteins of Bacillus thuringiensis show a high degree of specificity. In... more The insecticidal crystal proteins of Bacillus thuringiensis show a high degree of specificity. In vitro binding studies with several crystal proteins demonstrated a correlation between toxicity and binding to receptors of larval midgut epithelial cells. In order to study the domain-function relationships of the toxic fragment, hybrid crystal proteins based on CrylA(b) and CrylC were constructed. Two out of 11 hybrid proteins constructed exhibited insecticidal activity. Both displayed an insectidial spectrum similar to that of the parental crystal protein from which the C-terminal part of the toxic fragment originated. In addition, in vitro binding studies directly demonstrated the involvement of the C-terminal part of the toxic fragment in receptor binding. These results demonstrate that the C-terminal part of the toxic fragment determines specific receptor binding, which in turn determines, to a large extent, the insect specificity.
Journal of Invertebrate Pathology, 1992
The in vitro binding of four insecticidal crystal proteins (ICPs) from Bacillus thuringiensis to ... more The in vitro binding of four insecticidal crystal proteins (ICPs) from Bacillus thuringiensis to midgut tissue sections of two lepidopteran (Manduca sexta and Plutella xylostella) and one coleopteran (Leptinotarsa decemlineata) species was immunocytochemically analyzed. Monoclonal ...
Journal of Invertebrate Pathology, 1996
Binding of different Bacillus thuringiensis insecticidal crystal proteins (ICPs) to the midgut ep... more Binding of different Bacillus thuringiensis insecticidal crystal proteins (ICPs) to the midgut epithelium of Spodoptera frugiperda larvae was characterized by binding experiments with midgut tissue sections and isolated brush border membrane vesicles. Our results show that ICPs interact with the microvilli of epithelial cells of S. frugiperda in two different ways. The first is typical of highly toxic proteins (like CryIC and CryID); this interaction is saturable and specific. In contrast, some nontoxic proteins (like CryIAb) interact nonspecifically with the microvilli, since the binding of this toxin is not affected by the presence of high concentrations of homologous competitor. The CryIC toxin binds to two brush border proteins of 40 and 44 kDa and the CryIAb toxin binds to a single protein of 150 kDa. Immunological detection of ingested B. thuringiensis ICPs on gut sections of S. frugiperda larvae revealed that CryIC and CryID toxins bound along the epithelial brush border microvilli membrane. Binding of the nontoxic protein CryIAb was also observed in the epithelial brush border membrane of fed larvae, but it was extremely weak, implying that this type of interaction occurs also in vivo although it is not related to toxicity.
Physiological Entomology, 1982
ABSTRACT The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-... more ABSTRACT The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-h photophase) Leptinotarsa decemlineata Say adults was characterized by high performance liquid chromatography, which revealed that for short-day beetles ecdysterone and ecdysone are the major constituents of the free ecdysteroids, whereas in long-day beetles products co-migrating with 2-deoxyecdysone and 2-deoxyecdysterone are just as abundant. Ecdysteroids were detected (15 ng/ml) only in the haemolymph of females. Ligation experiments showed that the induction of the diapause syndrome by inactivation of the corpora allata or the brain not only resulted in the formation of the typical diapause proteins but also in an increase of the ecdysteroid activity. It is suggested that adult diapause in L.decemlineata is regulated by a multifactorial system.
Journal of Cellular Biochemistry, 1986
ACS Symposium Series, 1992
66th International Symposium on Crop Protection, 2014
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1986
Isolated ovaries of egg laying females synthesize and secrete three yolk proteins (two vitellogen... more Isolated ovaries of egg laying females synthesize and secrete three yolk proteins (two vitellogenins and chromoprotein 2). 2. The contribution of ovarian tissue to total yolk protein production is very small, the major site of synthesis of the three yolk proteins being the fat body. 3. There is a time lag between yolk protein synthesis by the fat body and yolk protein sequestration by the ovary. 4. In egg laying females, within 1 hr after the synthesis of both vitellogenins by the fat body, they appear in the oocytes as vitellins.
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1982
The 2 vitellogenins, VFPI and VFP2, are immunologically unrelated. 2. Both the viteUogenins are l... more The 2 vitellogenins, VFPI and VFP2, are immunologically unrelated. 2. Both the viteUogenins are lipo-and glycoproteins and consist of 4 and 6 polypeptides respectively. 3. The selective uptake of proteins (2 vitellogenins and 1 chromoprotein) into the oocyte, loads them with more negative charges, which is apparently the only change. 4. In prediapausing beetles, 3 immunologically unrelated proteins accumulate. One of these diapause proteins, DP1, also accumulates in larval and pupal hemolymph. 5. The diapause proteins are all glycoproteins, consisting of one single polypeptide.
Insect Biochemistry, 1984
The accessory reproductive glands (ARGs) of the male Colorado beetle secrete about 50 polypeptide... more The accessory reproductive glands (ARGs) of the male Colorado beetle secrete about 50 polypeptides with molecular weights varying between 15,000 and 650,000. Most of these polypeptides are synthesized by the gland itself. However, some secretory polypeptides originate from the fat body and are transported to the ARGs by the haemolymph. One of these extraglandular secretory polypeptides corresponds with one of the two subunits of the major haemolymph lipoprotein. The second subunit of this lipoprotein is not secreted, but is accumulated at the microvillar zone of the secretory cells.
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1986
Two vitellogenins and chromoprotein 2 are selectively accumulated by the oocyte and cannot be det... more Two vitellogenins and chromoprotein 2 are selectively accumulated by the oocyte and cannot be detected either in follicle cells or in the germarium. 2. At the start of their accumulation in terminal oocytes they are asymmetrically distributed. 3. Endocytosis of vitellogenin l starts somewhat later than the uptake of vitellogenin 2 and chromoprotein 2. 4. In follicle cells of young follicles, a protein (DLP), immunologically related to diapause protein l, is highly concentrated. 5. During vitellogenesis DLP is sequestered by the oocytes. 6. The protein rich globules in terminal oocytes contain the vitellins as well as chromoprotein 2 and DLP.
Journal of Insect Physiology, 1982
ABSTRACT
FEMS Microbiology Letters, 1989
Canadian Journal of Microbiology, 1996
Light microscopy was used to investigate the relation between toxicity, cytopathological effects,... more Light microscopy was used to investigate the relation between toxicity, cytopathological effects, and in vivo binding of Bacillus thuringiensis CrylA(b) and CrylE toxin proteins in larvae of Lymantria dispar, Choristoneura fumiferana, Actebia fennica, and Bombyx mori. These target insects were selected for their contrasting susceptibility to the two toxins. Lymantria dispar is susceptible to CrylA(b), B. mori is susceptible to CrylE, C. fumiferana is susceptible to both, and A. fennica is not susceptible to either. In the susceptible species, both toxins caused typical pathological changes in midgut epithelial cells, including disruption and shedding of the brush border membrane, vacuolization of the cytoplasm, and swelling of the cells and their nuclei, followed by disintegration and release of cytoplasmic content into the lumen. In the highly resistant A. fennica, no cell damage was observed, but the midguts of toxin-fed larvae had a shrunken appearance. Immunohistochemical staini...
Applied and Environmental Microbiology, 1993
According to the information supplied by the manufacturer of the biotinylation kit (RPN28), the c... more According to the information supplied by the manufacturer of the biotinylation kit (RPN28), the concentration of the N-hydroxysuccinimide-biotin ester was 0.46 mg/ml. However, recently we were told that this information was incorrect and that the actual concentration was 5 mg/ml. Therefore, the molar reaction ratios of biotin molecules to insecticidal crystal protein molecules in our paper should be multiplied by 10.9. (Note that these biotinylation ratios refer to the biotin-to-protein ratio during biotinylation [Anal. Biochem 195:214-219, 1991] rather than to the number of biotins actually attached.
Pest Management in Rice, 1990
During recent years several procedures have been developed to introduce foreign genes into plant ... more During recent years several procedures have been developed to introduce foreign genes into plant cells and to subsequently regenerate fertile plants. In combination with an increasing understanding of plant gene regulation and the processing and targeting of proteins this has provided the opportunity to obtain genetically modified plants. Traits of agronomic importance can be introduced into crop plants, one example being the engineering of plants resistant to insect attack through the expression of insecticidal proteins. At present the preferential source of such insecticidal proteins is the bacterium Bacillus thuringiensis (B.t.).
European Journal of Biochemistry, 1997
We report the purification, cloning and characterization of an aminopeptidase N from the midgut e... more We report the purification, cloning and characterization of an aminopeptidase N from the midgut epithelium of Manduca sexta that binds Cry1 Ab5, an insecticidal crystal protein [ICP] from Bacillus fhuringiensis. Sequence information derived from this M. sexta aminopeptidase N was used for the cloning o f an aminopeptidase N from the midgut brush-border membrane of Plutrlla xylostella, an insect species of which some populations acquired resistance against Cry1 Ab5. Affinity chromatography on a CrylAb5 matrix was used to isolate a 220-kDa glycoprotein from the larval midgut of the lepidopteran M. sexta. On ligand blots the purified 120-kDa protein discriminates between the lepidopteran-specific Cry1 Ab5 and the coleopteran-specific Cry3A d-endotoxin. Internal amino acid sequences from the 120-kDa protein were used for the design of degenerate oligonucleotides. From a nested PCR with M. sexta midgut cDNA as template, a DNA fragment was obtained which shows similarity to prokaryotic and eukaryotic aminopeptidase N genes. This PCR fragment was used to screen cDNA libraries of larval midguts from M. sexta and P. xylostella. From the M. sextu midgut cDNA library a 2973-bp nucleotide sequence was cloned. The ORF of the sequence encodes a 942-residue aminopeptidase N (M. sexta Apn2) containing two hydrophobic regions. The NH,-terminal hydrophobic region corresponds to a secretory signal sequence and the COOH-terminal hydrophobic region is typical of glycosylphosphatidylinositol (glycosyl-PtdTns)-anchored proteins. Low-stringency hybridization of the tl xylostella midgut cDNA library with M. sexta u p 2 probes enabled the isolation of a 3118-bp sequence with an ORF encoding a 946-residue preproprotein. This aminopeptidase N (P xylostella Apnl) displays 61 % amino acid identity to M. sexta Apn2 and contains a COOH-terminal signal peptide for glycosyl-PtdIns anchor addition. Both M. sexta Apn2 and P. .xy'lostella Apnl contain four Cys residues, which are highly conserved among eukaryotic aminopeptidase N molecules. Treatment of Sf9 cells expressing the P xylostellu apnl gene with PtdIns-specific phospholipase C demonstrated that tl xylostella Apnl is attached to the insect cell tnembrane by a glycosyl-Ptdlns anchor.
Proceedings of the National Academy of Sciences, 1991
The biochemical mechanism for resistance to Bacillus thuringiensis crystal proteins was studied i... more The biochemical mechanism for resistance to Bacillus thuringiensis crystal proteins was studied in a field population of diamondback moths (Plutella xylostella) with a reduced susceptibility to the bioinsecticidal spray. The toxicity and binding characteristics of three crystal proteins [CryIA(b), CryIB, and CryIC] were compared between the field population and a laboratory strain. The field population proved resistant (greater than 200-fold compared with the laboratory strain) to CryIA(b), one of the crystal proteins in the insecticidal formulation. Binding studies showed that the two strains differ in a membrane receptor that recognizes CryIA(b). This crystal protein did not bind to the brush-border membrane of the midgut epithelial cells of the field population, either because of strongly reduced binding affinity or because of the complete absence of the receptor molecule. Both strains proved fully susceptible to the CryIB and CryIC crystal proteins, which were not present in the...
Molecular Microbiology, 1991
The insecticidal crystal proteins of Bacillus thuringiensis show a high degree of specificity. In... more The insecticidal crystal proteins of Bacillus thuringiensis show a high degree of specificity. In vitro binding studies with several crystal proteins demonstrated a correlation between toxicity and binding to receptors of larval midgut epithelial cells. In order to study the domain-function relationships of the toxic fragment, hybrid crystal proteins based on CrylA(b) and CrylC were constructed. Two out of 11 hybrid proteins constructed exhibited insecticidal activity. Both displayed an insectidial spectrum similar to that of the parental crystal protein from which the C-terminal part of the toxic fragment originated. In addition, in vitro binding studies directly demonstrated the involvement of the C-terminal part of the toxic fragment in receptor binding. These results demonstrate that the C-terminal part of the toxic fragment determines specific receptor binding, which in turn determines, to a large extent, the insect specificity.
Journal of Invertebrate Pathology, 1992
The in vitro binding of four insecticidal crystal proteins (ICPs) from Bacillus thuringiensis to ... more The in vitro binding of four insecticidal crystal proteins (ICPs) from Bacillus thuringiensis to midgut tissue sections of two lepidopteran (Manduca sexta and Plutella xylostella) and one coleopteran (Leptinotarsa decemlineata) species was immunocytochemically analyzed. Monoclonal ...
Journal of Invertebrate Pathology, 1996
Binding of different Bacillus thuringiensis insecticidal crystal proteins (ICPs) to the midgut ep... more Binding of different Bacillus thuringiensis insecticidal crystal proteins (ICPs) to the midgut epithelium of Spodoptera frugiperda larvae was characterized by binding experiments with midgut tissue sections and isolated brush border membrane vesicles. Our results show that ICPs interact with the microvilli of epithelial cells of S. frugiperda in two different ways. The first is typical of highly toxic proteins (like CryIC and CryID); this interaction is saturable and specific. In contrast, some nontoxic proteins (like CryIAb) interact nonspecifically with the microvilli, since the binding of this toxin is not affected by the presence of high concentrations of homologous competitor. The CryIC toxin binds to two brush border proteins of 40 and 44 kDa and the CryIAb toxin binds to a single protein of 150 kDa. Immunological detection of ingested B. thuringiensis ICPs on gut sections of S. frugiperda larvae revealed that CryIC and CryID toxins bound along the epithelial brush border microvilli membrane. Binding of the nontoxic protein CryIAb was also observed in the epithelial brush border membrane of fed larvae, but it was extremely weak, implying that this type of interaction occurs also in vivo although it is not related to toxicity.