Plant α‐amylase inhibitors and their interaction with insect α‐amylases

Insect pests and pathogens (fungi, bacteria and viruses) are responsible for severe crop losses. Insects feed directly on the plant tissues, while the pathogens lead to damage or death of the plant. Plants have evolved a certain degree of resistance through the production of defence compounds, which may be aproteic, e.g. antibiotics, alkaloids, terpenes, cyanogenic glucosides or proteic, e.g. chitinases, β-1,3-glucanases, lectins, arcelins, vicilins, systemins and enzyme inhibitors. The enzyme inhibitors impede digestion through their action on insect gut digestive α-amylases and proteinases, which play a key role in the digestion of plant starch and proteins. The natural defences of crop plants may be improved through the use of transgenic technology. Current research in the area focuses particularly on weevils as these are highly dependent on starch for their energy supply. Six different α-amylase inhibitor classes, lectin-like, knottin-like, cereal-type, Kunitz-like, γ-purothionin-like and thaumatin-like could be used in pest control. These classes of inhibitors show remarkable structural variety leading to different modes of inhibition and different specificity profiles against diverse α-amylases. Specificity of inhibition is an important issue as the introduced inhibitor must not adversely affect the plant's own α-amylases, nor the nutritional value of the crop. Of particular interest are some bifunctional inhibitors with additional favourable properties, such as proteinase inhibitory activity or chitinase activity. The area has benefited from the recent determination of many structures of α-amylases, inhibitors and complexes. These structures highlight the remarkable variety in structural modes of α-amylase inhibition. The continuing discovery of new classes of α-amylase inhibitor ensures that exciting discoveries remain to be made. In this review, we summarize existing knowledge of insect α-amylases, plant α-amylase inhibitors and their interaction. Positive results recently obtained for transgenic plants and future prospects in the area are reviewed.

[1]  Robert Huber,et al.  A novel strategy for inhibition of α-amylases: yellow meal worm α-amylase in complex with the Ragi bifunctional inhibitor at 2.5 å resolution , 1998 .

[2]  D. Rigden,et al.  Purification, biochemical characterisation and partial primary structure of a new α-amylase inhibitor from Secale cereale (rye) , 2000 .

[3]  T. Morimoto,et al.  Tertiary and quaternary structures of 0.19 alpha-amylase inhibitor from wheat kernel determined by X-ray analysis at 2.06 A resolution. , 1997, Biochemistry.

[4]  N. Juge,et al.  Specific inhibition of barley α‐amylase 2 by barley α‐amylase/subtilisin inhibitor depends on charge interactions and can be conferred to isozyme 1 by mutation , 2000 .

[5]  C. Betzel,et al.  Synchrotron X-ray data collection and restrained least-squares refinement of the crystal structure of proteinase K at 1.5 A resolution. , 1988, Acta Crystallographica Section B Structural Science.

[6]  B. Cornelissen,et al.  A tobacco mosaic virus-induced tobacco protein is homologous to the sweet-tasting protein thaumatin , 1986, Nature.

[7]  P. Shewry,et al.  Purification and characterization of an insect alpha-amylase inhibitor/endochitinase from seeds of Job's Tears (Coix lachryma-jobi). , 1989, Biochimica et biophysica acta.

[8]  The inhibition of human salivary alpha-amylase by type II alpha-amylase inhibitor from Triticum aestivum is competitive, slow and tight-binding. , 1995, Journal of enzyme inhibition.

[9]  M. Chrispeels,et al.  Molecular cloning of bruchid (Zabrotes subfasciatus) alpha-amylase cDNA and interactions of the expressed enzyme with bean amylase inhibitors. , 1997, Insect biochemistry and molecular biology.

[10]  G Neshich,et al.  Analysis of structural and physico-chemical parameters involved in the specificity of binding between alpha-amylases and their inhibitors. , 2000, Protein engineering.

[11]  T. Arakawa,et al.  A Chimera-Like α-Amylase Inhibitor Suggesting the Evolution of Phaseolus vulgaris α-Amylase Inhibitor , 2000 .

[12]  F. Garcia-Maroto,et al.  Cloning of cDNA and chromosomal location of genes encoding the three types of subunits of the wheat tetrameric inhibitor of insect α-amylase , 1990, Plant Molecular Biology.

[13]  T. Higgins,et al.  Bean alpha-amylase inhibitor 1 in transgenic peas (Pisum sativum) provides complete protection from pea weevil (Bruchus pisorum) under field conditions. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[14]  T. Singh,et al.  Stability and kinetics of a bifunctional amylase/trypsin inhibitor. , 1993, Biochimica et biophysica acta.

[15]  C. Ryan.  Protease Inhibitors in Plants: Genes for Improving Defenses Against Insects and Pathogens , 1990 .

[16]  C. Park,et al.  Mapping function to structure in a channel-blocking peptide: electrostatic mutants of charybdotoxin. , 1992, Biochemistry.

[17]  F. White,et al.  The corn inhibitor of activated Hageman factor: purification and properties of two recombinant forms of the protein. , 1998, Protein expression and purification.

[18]  V. Silano,et al.  Further characterization studies of the alpha-amylase protein inhibitor of gel electrophoretic mobility 0.19 from the wheat kernel. , 1976, Biochimica et biophysica acta.

[19]  J. Jacobsen,et al.  Regulation of synthesis and transport of secreted proteins in cereal aleurone. , 1991, International review of cytology.

[20]  A. Blanco-Labra,et al.  PURIFICATION AND CHARACTERIZATION OF AN α‐AMYLASE INHIBITOR FROM MAISE (ZEA MAIZE) , 1981 .

[21]  P. Rougé,et al.  A plant-seed inhibitor of two classes of alpha-amylases: X-ray analysis of Tenebrio molitor larvae alpha-amylase in complex with the bean Phaseolus vulgaris inhibitor. , 1999, Acta crystallographica. Section D, Biological crystallography.

[22]  S. Pongor,et al.  Solution Structure of the Major α-Amylase Inhibitor of the Crop Plant Amaranth* , 1999, The Journal of Biological Chemistry.

[23]  T. Mirkov,et al.  Evolutionary relationships among proteins in the phytohemagglutinin-arcelin-α-amylase inhibitor family of the common bean and its relatives , 1994, Plant Molecular Biology.

[24]  W. Saenger,et al.  The three‐dimensional structure of the bifunctional proteinase k/α‐amylase inhibitor from wheat (PKI3) at 2.5 Å resolution , 1991 .

[25]  T. Arakawa,et al.  Structural Characterization of an α-Amylase Inhibitor from a Wild Common Bean (Phaseolus vulgaris): Insight into the Common Structural Features of Leguminous α-Amylase Inhibitors , 1997 .

[26]  B. Svensson,et al.  Arginine is essential for the α-amylase inhibitory activity of the α-amylase/subtilisin inhibitor (BASI) from barley seeds , 1993 .

[27]  W. K. Roberts,et al.  A new family of plant antifungal proteins. , 1991, Molecular plant-microbe interactions : MPMI.

[28]  M. Chrispeels,et al.  A lectin gene encodes the alpha-amylase inhibitor of the common bean. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Hee-Seob Lee,et al.  Comparative Study of the Inhibition of α-Glucosidase, α-Amylase, and Cyclomaltodextrin Glucanosyltransferase by Acarbose, Isoacarbose, and Acarviosine–Glucose , 1999 .

[30]  J. Robertus,et al.  Structure of ricin B‐chain at 2.5 Å resolution , 1991, Proteins.

[31]  J. Kawabata,et al.  α-Amylase Inhibitors from Roselle (Hibiscus sabdariffa Linn.) Tea , 2000, Bioscience, biotechnology, and biochemistry.

[32]  P. Shewry,et al.  Characterization of homologous inhibitors of trypsin and α-amylase from seeds of rye (Secale cereale L.) , 1987 .

[33]  T. Singh,et al.  Substrate−Inhibitor Interactions in the Kinetics of α-Amylase Inhibition by Ragi α-amylase/Trypsin Inhibitor (RATI) and Its Various N-Terminal Fragments† , 2001 .

[34]  M. D. O'donnell,et al.  Differential serum amylase determination by use of an inhibitor, and design of a routine procedure. , 1977, Clinical chemistry.

[35]  Hughes,et al.  Defense proteins from seed of Cassia fistula include a lipid transfer protein homologue and a protease inhibitory plant defensin. , 2000, Plant science : an international journal of experimental plant biology.

[36]  R. Huber,et al.  Determination of the three-dimensional structure of the bifunctional alpha-amylase/trypsin inhibitor from ragi seeds by NMR spectroscopy. , 1995, Biochemistry.

[37]  A. Valencia-Jiménez,et al.  Digestive amylase from the larger grain borer, Prostephanus truncatus Horn. , 2000, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[38]  M. Sinnott,et al.  Catalytic mechanism of enzymic glycosyl transfer , 1990 .

[39]  S. Pongor,et al.  A novel alpha-amylase inhibitor from amaranth (Amaranthus hypocondriacus) seeds. , 1994, The Journal of biological chemistry.

[40]  David C. Jones,et al.  CATH--a hierarchic classification of protein domain structures. , 1997, Structure.

[41]  J M Thornton,et al.  1H NMR structure of an antifungal γ‐thionin protein SIα1: Similarity to scorpion toxins , 1998, Proteins.

[42]  C. Bloch,et al.  A new family of small (5 kDa) protein inhibitors of insect α‐amylases from seeds or sorghum (Sorghum bicolor (L) Moench) have sequence homologies with wheat γ‐purothionins , 1991 .

[43]  R. Sweet,et al.  Crystal structure of thermostable alpha-amylase from Bacillus licheniformis refined at 1.7 A resolution. , 1997, Molecules and cells.

[44]  G. Ossa,et al.  Alpha-amylases of the coffee berry borer (Hypothenemus hampei) and their inhibition by two plant amylase inhibitors. , 2000, Insect biochemistry and molecular biology.

[45]  F. Lajolo,et al.  Amylase inhibitors in Phaseolus vulgaris beans , 1991 .

[46]  Acarbose ‐ ein neues Wirkprinzip in der Diabetestherapie , 1994 .

[47]  S. Hizukuri,et al.  STUDIES ON UDPG: ALPHA-1,4-GLUCAN ALPHA-4-GLUCOSYLTRANSFERASE. VII. CONVERSION OF THE ENZYME FROM GLUCOSE-6-PHOSPHATE-DEPENDENT TO INDEPENDENT FORM IN LIVER. , 1964, Biochemistry.

[48]  G. García-Casado,et al.  Role of complex asparagine-linked glycans in the allergenicity of plant glycoproteins. , 1996, Glycobiology.

[49]  A. Blanco-Labra,et al.  A possible function for thaumatin and a TMV-induced protein suggested by homology to a maize inhibitor , 1987, Nature.

[50]  F. Garcia-Maroto,et al.  Site-directed mutagenesis and expression inEscherichia coli of WMAI-1, a wheat monomeric inhibitor of insect α-amylase , 1991, Plant Molecular Biology.

[51]  T. Arakawa,et al.  Complete Sequence, Subunit Structure, and Complexes with Pancreatic α-Amylase of an α-Amylase Inhibitor from Phaseolus vulgaris White Kidney Beans , 1996 .

[52]  L. Nadeau,et al.  A new rapid immunoinhibition pancreatic amylase assay: diagnostic value for pancreatitis. , 1994, Clinical biochemistry.

[53]  H. Nakatani,et al.  In vitro action of human and porcine α-amylases on cyclomalto-oligosaccharides , 1990 .

[54]  C. Brownlee,et al.  Fungal Membrane Responses Induced by Plant Defensins and Thionins* , 1996, The Journal of Biological Chemistry.

[55]  F. Campos,et al.  The complete amino acid sequence of the bifunctional α‐amylase/trypsin inhibitor from seeds of ragi (Indian finger millet, Eleusine coracana Gaertn.) , 1983 .

[56]  C. Selitrennikoff,et al.  Zeamatin Inhibits Trypsin and α-Amylase Activities , 2001, Applied and Environmental Microbiology.

[57]  T. Higgins,et al.  Bean alpha-amylase inhibitor 1 in transgenic peas (Pisum sativum) provides complete protection from pea weevil (Bruchus pisorum) under field conditions. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[58]  A G Murzin,et al.  SCOP: a structural classification of proteins database for the investigation of sequences and structures. , 1995, Journal of molecular biology.

[59]  G. Pearce,et al.  Systemin: a polypeptide signal for plant defensive genes. , 1998, Annual review of cell and developmental biology.

[60]  Ian Denholm,et al.  Insect-resistant transgenic plants , 1998 .

[61]  C. Potvin,et al.  Several Thaumatin-Like Proteins Bind to β-1,3-Glucans , 1998 .

[62]  B. Svensson,et al.  Improved Activity and Modulated Action Pattern Obtained by Random Mutagenesis at the Fourth β-α Loop Involved in Substrate Binding to the Catalytic (β/α)8-Barrel Domain of Barley α-Amylase 1* , 1997, The Journal of Biological Chemistry.

[63]  R. J. Harley,et al.  Recording System for Monitoring Automaticity of Heart Cells in Culture , 1971, Nature.

[64]  C. Orengo,et al.  Alpha plus beta folds revisited: some favoured motifs. , 1993, Structure.

[65]  T. A. Valueva,et al.  Enzymatic oxidation of the bifunctional wheat inhibitor of subtilisin and endogenous α‐amylase , 1993 .

[66]  M. Calero,et al.  Primary structure of omega-hordothionin, a member of a novel family of thionins from barley endosperm, and its inhibition of protein synthesis in eukaryotic and prokaryotic cell-free systems. , 1996, European journal of biochemistry.

[67]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[68]  J. J. Marshall,et al.  Purification and properties of phaseolamin, an inhibitor of alpha-amylase, from the kidney bean, Phaseolus vulgaris. , 1975, The Journal of biological chemistry.

[69]  A. Monzingo,et al.  The crystal structure of the antifungal protein zeamatin, a member of the thaumatin-like, PR-5 protein family , 1996, Nature Structural Biology.

[70]  W. K. Roberts,et al.  Zeamatin, an antifungal protein from maize with membrane-permeabilizing activity , 1990 .

[71]  K. Okuyama,et al.  Crystal structure of Thermoactinomyces vulgaris R-47 alpha-amylase II (TVAII) hydrolyzing cyclodextrins and pullulan at 2.6 A resolution. , 1999, Journal of molecular biology.

[72]  C. Bompard-Gilles,et al.  Characterization and functional properties of the α-amylase inhibitor (α-AI) from kidney bean (Phaseolus vulgaris) seeds , 1997 .

[73]  K. Ohtsubo,et al.  The amino acid sequence of a 20 kDa bifunctional subtilisin/α‐amylase inhibitor from brain of rice (Oryza saliva L.) seeds , 1992 .

[74]  B. Vallee,et al.  Metal content of alpha-amylases of various origins. , 1959, The Journal of biological chemistry.

[75]  É. Várallyay,et al.  Arg-27, Arg-127 and Arg-155 in the beta-trefoil protein barley alpha-amylase/subtilisin inhibitor are interface residues in the complex with barley alpha-amylase 2. , 1995, The Biochemical journal.

[76]  M. F. Grossi-de-Sá,et al.  Do legume storage proteins play a role in defending seeds against bruchids? , 2000, Plant physiology.

[77]  T. Higgins,et al.  Expression of the insecticidal bean alpha-amylase inhibitor transgene has minimal detrimental effect on the nutritional value of peas fed to rats at 30% of the diet. , 1999, The Journal of nutrition.

[78]  P. Hasegawa,et al.  Antifungal activity of tobacco osmotin has specificity and involves plasma membrane permeabilization , 1996 .

[79]  R. Shibata,et al.  CM3, one of the wheat alpha-amylase inhibitor subunits, and binding of IgE in sera from Japanese with atopic dermatitis related to wheat. , 2000, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[80]  Š. Janeček,et al.  Relationship of sequence and structure to specificity in the α-amylase family of enzymes , 2001 .

[81]  B. Svensson,et al.  Stopped‐flow kinetic studies of the reaction of barley α‐amylase/subtilisin inhibitor and the high pI barley α‐amylase , 1995 .

[82]  M. Ishimoto,et al.  Insecticidal activity of an α-amylase inhibitor-like protein resembling a putative precursor of α-amylase inhibitor in the common bean, Phaseolus vulgaris L. , 1999 .

[83]  G. Reeck,et al.  alpha-Amylase inhibitors from wheat: amino acid sequences and patterns of inhibition of insect and human alpha-amylases. , 1996, Insect biochemistry and molecular biology.

[84]  F. Payan,et al.  Carbohydrate binding sites in a pancreatic α‐amylase‐substrate complex, derived from X‐ray structure analysis at 2.1 Å resolution , 1995, Protein science : a publication of the Protein Society.

[85]  H. Kaji,et al.  Overexpression in Escherichia coli of chemically synthesized gene for active 0.19 alpha-amylase inhibitor from wheat kernel. , 1997, Journal of biochemistry.

[86]  J G Bishop,et al.  Rapid evolution in plant chitinases: molecular targets of selection in plant-pathogen coevolution. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[87]  B. Svensson,et al.  Barley alpha-amylase bound to its endogenous protein inhibitor BASI: crystal structure of the complex at 1.9 A resolution. , 1998, Structure.

[88]  E. Titarenko,et al.  cDNA cloning, biochemical characterization and inhibition by plant inhibitors of the alpha-amylases of the Western corn rootworm, Diabrotica virgifera virgifera. , 2000, Insect biochemistry and molecular biology.

[89]  M. Ishimoto,et al.  Protective Mechanism of the Mexican Bean Weevil against High Levels of [alpha]-Amylase Inhibitor in the Common Bean , 1996, Plant physiology.

[90]  D. Boulter,et al.  A novel mechanism of insect resistance engineered into tobacco , 1987, Nature.

[91]  P. Brick,et al.  Crystal structure of a Kunitz-type trypsin inhibitor from Erythrina caffra seeds. , 1991, Journal of molecular biology.

[92]  C. López-Otín,et al.  A major baker's asthma allergen from rye flour is considerably more active than its barley counterpart , 1995, FEBS letters.

[93]  M. Ishimoto,et al.  Growth Inhibitory Effects of an α-Amylase Inhibitor from the Kidney Bean, Phaseolus vulgaris (L.) on Three Species of Bruchids (Coleoptera : Bruchidae) , 1989 .

[94]  L. Pedersen,et al.  Structural determinants of the bifunctional corn Hageman factor inhibitor: x-ray crystal structure at 1.95 A resolution. , 1998, Biochemistry.

[95]  C. Woloshuk,et al.  Inhibition of growth of Aspergillus flavus and fungal alpha-amylases by a lectin-like protein from Lablab purpureus. , 2001, Molecular plant-microbe interactions : MPMI.

[96]  S. Gollasch,et al.  Bean [alpha]-Amylase Inhibitor Confers Resistance to the Pea Weevil (Bruchus pisorum) in Transgenic Peas (Pisum sativum L.) , 1995, Plant physiology.

[97]  D. Boulter,et al.  Potential of plant-derived genes in the genetic manipulation of crops for insect resistance , 1992 .

[98]  S. Jacobsen,et al.  Two antifungal thaumatin‐like proteins from barley grain , 1991, FEBS letters.

[99]  A. Mazur,et al.  The catalytic mechanism of alpha-amylases based upon enzyme crystal structures and model building calculations. , 1994, Biochemical and biophysical research communications.

[100]  M. Chrispeels,et al.  A putative precursor protein in the evolution of the bean α-amylase inhibitor , 1996 .

[101]  V. Buonocore,et al.  Interaction of Tenebrio molitor L. α‐amylase with a wheat flour protein inhibitor , 1976 .

[102]  W. Terra,et al.  Apocrine secretion of amylase and exocytosis of trypsin along the midgut of Tenebrio molitor larvae. , 2001, Journal of insect physiology.

[103]  S. Koyama,et al.  Kinetic Studies on the Hydrolyses of α-, β-, and γ-Cyclodextrins by Taka-amylase A , 1974 .

[104]  C. López-Otín,et al.  Rye inhibitors of animal alpha-amylases show different specificities, aggregative properties and IgE-binding capacities than their homologues from wheat and barley. , 1994, European journal of biochemistry.

[105]  H. Yamaguchi.  Isolation and characterization of the subunits of a heat-labile alpha-amylase inhibitor from Phaseolus vulgaris white kidney bean. , 1993, Bioscience, biotechnology, and biochemistry.

[106]  J. Mundy,et al.  Barley α-amylase/subtilisin inhibitor. I. Isolation and characterization , 1983 .

[107]  G. Reeck,et al.  α-amylases from three species of stored grain coleoptera and their inhibition by wheat and corn proteinaceous inhibitors☆ , 1992 .

[108]  S. Wodak,et al.  Solution structure of the main α‐amylase inhibitor from amaranth seeds , 2001 .

[109]  V. Silano,et al.  Wheat protein inhibitors of α-amylase , 1977 .

[110]  M. Richardson,et al.  The complete amino acid sequence of a major wheat protein inhibitor of α-amylase , 1981 .

[111]  Lynn J. Frewer,et al.  ASSESSING AND STRUCTURING ATTITUDES TOWARD THE USE OF GENE TECHNOLOGY IN FOOD-PRODUCTION - THE ROLE OF PERCEIVED ETHICAL OBLIGATION , 1995 .

[112]  J. Whitaker,et al.  STRUCTURAL FEATURES OF RED KIDNEY BEAN á-AMYLASE INHIBITOR IMPORTANT IN BINDING WITH á-AMYLASE , 1984 .

[113]  P. Thibault,et al.  Post‐translational processing of two α‐amylase inhibitors and an arcelin from the common bean, Phaseolus vulgaris , 1999 .

[114]  C. Silva,et al.  Digestion in larvae of Callosobruchus maculatus and Zabrotes subfasciatus (Coleoptera: Bruchidae) with emphasis on α-amylases and oligosaccharidases , 1999 .

[115]  A. Giri,et al.  Amylase inhibitors of pigeonpea (Cajanus cajan) seeds. , 1998, Phytochemistry.

[116]  R. Huber,et al.  Specific inhibition of insect α-amylases: yellow meal worm α-amylase in complex with the Amaranth α-amylase inhibitor at 2.0 Å resolution , 1999 .

[117]  G. Salcedo,et al.  New dimeric inhibitor of heterologous alpha-amylases encoded by a duplicated gene in the short arm of chromosome 3B of wheat (Triticum aestivum L.). , 1989, European journal of biochemistry.

[118]  J. Gatehouse,et al.  Identifying proteins with insecticidal activity: use of encoding genes to produce insect‐resistant transgenic crops , 1998 .

[119]  T. Higgins,et al.  Genetic engineering with α-amylase inhibitors makes seeds resistant to bruchids , 1998, Seed Science Research.

[120]  B. Svensson,et al.  Molecular structure of a barley alpha-amylase-inhibitor complex: implications for starch binding and catalysis. , 1998, Journal of molecular biology.

[121]  P. Rougé,et al.  Mechanism of porcine pancreatic alpha-amylase inhibition of amylose and maltopentaose hydrolysis by kidney bean (Phaseolus vulgaris) inhibitor and comparison with that by acarbose. , 1999, European journal of biochemistry.

[122]  R. Huber,et al.  Crystal structure of calcium-depleted Bacillus licheniformis alpha-amylase at 2.2 A resolution. , 1995, Journal of molecular biology.

[123]  T. Higgins,et al.  Transgenic Pea Seeds Expressing the α-Amylase Inhibitor of the Common Bean are Resistant to Bruchid Beetles , 1994, Bio/Technology.

[124]  R. Glockshuber,et al.  RBI, a one‐domain α‐amylase/trypsin inhibitor with completely independent binding sites , 1996 .

[125]  E. Dimagno,et al.  Partially purified white bean amylase inhibitor reduces starch digestion in vitro and inactivates intraduodenal amylase in humans. , 1985, Gastroenterology.

[126]  S. Larson,et al.  Refined Molecular Structure of Pig Pancreatic α-Amylase at 2·1 Å Resolution , 1994 .

[127]  A. Konarev.  Interaction of insect digestive enzymes with plant protein inhibitors and host-parasite coevolution , 2004, Euphytica.

[128]  C. Bompard-Gilles,et al.  Substrate mimicry in the active center of a mammalian alpha-amylase: structural analysis of an enzyme-inhibitor complex. , 1996, Structure.

[129]  P. Rougé,et al.  Molecular basis for the specific binding of different α-amylase inhibitors from Phaseolus vulgaris seeds to the active site of α-amylase , 2000 .

[130]  R. Huber,et al.  Structural basis of the endoproteinase-protein inhibitor interaction. , 2000, Biochimica et biophysica acta.

[131]  C. Betzel,et al.  Crystallization of the bifunctional proteinase/amylase inhibitor PKI‐3 and of its complex with proteinase K , 1986, FEBS letters.

[132]  A. Cohen,et al.  Partial characterization of α-amylase in the salivary glands of Lygus hesperus and L. lineolaris. , 2000 .

[133]  M. Ishimoto,et al.  Molecular characterization of a bean α-amylase inhibitor that inhibits the α-amylase of the Mexican bean weevil Zabrotes subfasciatus , 1997, Planta.

[134]  R. Adar,et al.  C-terminal Post-translational Proteolysis of Plant Lectins and Their Recombinant Forms Expressed in Escherichia coli , 1995, The Journal of Biological Chemistry.

[135]  H. Kamasaka,et al.  Rice bifunctional alpha-amylase/subtilisin inhibitor: characterization, localization, and changes in developing and germinating seeds. , 1998, Bioscience, biotechnology, and biochemistry.

[136]  J. Whitaker,et al.  Some aspects of the mechanism of complexation of red kidney bean alpha-amylase inhibitor and alpha-amylase. , 1984, Biochemistry.

[137]  M. Giband,et al.  Transgenic Plants for Insect Resistance , 1998 .

[138]  J. Pueyo,et al.  Activation of Bean (Phaseolus vulgaris) [alpha]-Amylase Inhibitor Requires Proteolytic Processing of the Proprotein , 1993, Plant physiology.

[139]  C. Cambillau,et al.  Crystal structure of pig pancreatic alpha-amylase isoenzyme II, in complex with the carbohydrate inhibitor acarbose. , 1996, European journal of biochemistry.

[140]  B. Svensson.  Protein engineering in the α-amylase family: catalytic mechanism, substrate specificity, and stability , 1994, Plant Molecular Biology.

[141]  Y. Moreau,et al.  Mechanism of porcine pancreatic alpha-amylase. Inhibition of amylose and maltopentaose hydrolysis by alpha-, beta- and gamma-cyclodextrins. , 2001, European journal of biochemistry.

[142]  R. Huber,et al.  Crystal structure of yellow meal worm alpha-amylase at 1.64 A resolution. , 1998, Journal of molecular biology.

[143]  R. Huber,et al.  Carbohydrate and protein-based inhibitors of porcine pancreatic alpha-amylase: structure analysis and comparison of their binding characteristics. , 1996, Journal of molecular biology.

[144]  M. Ishimoto,et al.  Bruchid resistance of transgenic azuki bean expressing seed α‐amylase inhibitor of common bean , 1996 .

[145]  H. Matsubara,et al.  Complete amino acid sequence of an α-amylase inhibitor in wheat kernel , 1983 .

[146]  D. Rigden,et al.  Activity of wheat α‐amylase inhibitors towards bruchid α‐amylases and structural explanation of observed specificities , 2000 .

[147]  H. Matsubara,et al.  Crystallization and preliminary X-ray diffraction studies of the alpha-amylase inhibitor coded 0.19 from wheat kernel. , 1994, Journal of biochemistry.

[148]  W. Terra,et al.  Digestion of legume starch granules by larvae of Zabrotes subfasciatus (Coleoptera: bruchidae) and the induction of alpha-amylases in response to different diets. , 2001, Insect biochemistry and molecular biology.

[149]  P. Sanders,et al.  Inheritance of Functional Foreign Genes in Plants , 1984, Science.

[150]  D. Fairlie,et al.  Conformational homogeneity in molecular recognition by proteolytic enzymes , 1999, Journal of molecular recognition : JMR.

[151]  F. Campos,et al.  Resolution and partial characterization of proteinases and α-amylases from midguts of larvae of the bruchid beetle Callosobruchus maculatus (F.) , 1989 .

[152]  B. Henrissat,et al.  Crystal structures of human pancreatic alpha-amylase in complex with carbohydrate and proteinaceous inhibitors. , 2000, The Biochemical journal.

[153]  J Otlewski,et al.  The refined 2.0 Å X‐ray crystal structure of the complex formed between bovine β‐trypsin and CMTI‐I, a trypsin inhibitor from squash seeds (Cucurbita maxima) Topological similarity of the squash seed inhibitors with the carboxypeptidase A inhibitor from potatoes , 1989, FEBS letters.