Richard Guillory - Academia.edu (original) (raw)
Papers by Richard Guillory
Biochem Biophys Res Commun, 1985
The purified mitochondrial NADH dehydrogenase enzyme has been shown to catalyze a rapid [4B-3H] N... more The purified mitochondrial NADH dehydrogenase enzyme has been shown to catalyze a rapid [4B-3H] NADH-H20 exchange reaction. When the enzyme is subjected to a single freeze-thaw cycle there is a complete loss of NADH dehydrogenation without a measurable decrease in the [4B-3H] NADH-H20 exchange. Complete loss of the [4B-3H] NADH-H20 exchange follows brief exposure to ultraviolet photoirradiation. The differential sensitivity of the water exchange reaction and the dehydrogenase activity suggests a direct involvement of the enzymes flavin cofactor in the catalysis of the [4B-3H] NADH-H20 exchange. Arylazido-S-alanyl NAD+ (A3'-0-{3-[N-4-azido-2-nitrophenyl)amino] propionyl}NAD+) is shown to be a potent photodependent inhibitor of the [4B-3H] NADH-H20 exchange activity following photoirradiation with visible light. This is consistent with the observed photodependent inhibition of the dehydrogenase activity by this photoprobe (Chen, S. and Guillory, R.J. (1981) J. Biol. Chem. 256, 8318-8323). The abbreviations used are: arylazido-S-alanyl NAD+, A3'-O-{3-[N-(4-azido-2nitrophenyl)amino]propionyl)NAD+, AcPyAD+, 3-acetyl pyridine adenine dinucleotide.
Proceedings of the National Academy of Sciences of the United States of America, 1994
Biochemistry Usa, 1997
The leukocyte iodonitrotetrazolium violet (INT) reductase activity of disrupted bovine polymorpho... more The leukocyte iodonitrotetrazolium violet (INT) reductase activity of disrupted bovine polymorphonuclear neutrophils is closely associated with the activation of the O 2-generating NADPH oxidase in a cell-free system. It is dependent upon NADPH, cytosolic factors, and amphiphiles (such as arachidonate), the same factors required for O 2-generation. Both O 2-generation and INT reductase activity are inhibited by phenylarsine oxide, an inhibitor of the activation of the NADPH oxidase [
Molecular Structure, Function, and Assembly of the ATP Synthases, 1989
Current Topics in Bioenergetics, 1979
Proceedings of the National Academy of Sciences of the United States of America, 1959
Biochimica et biophysica acta, Jan 26, 1964
I. The antibiotic Dio-9 inhibits the acceptor-controUed respiration of intact rat-liver mitochond... more I. The antibiotic Dio-9 inhibits the acceptor-controUed respiration of intact rat-liver mitochondria in a phosphate-containing medium. Essentially the same effects have been found with glutamate, fl-hydroxybutyrate, a-ketoglutarate, succinate and the ascorbate-tetramethyl-p-phenylenediamine system as substrate. 2. Inhibition of respiration by Dio-9 in a phosphate-containing system cannot be reversed by 2,4-dinitrophenol. However, if 2,4-dinitrophenol is added in uncoupling concentrations (o.I raM) prior to the antibiotic there is no inhibition of respiration by Dio-9. 3. The inhibition of respiration by Dio-9 is preceded by a short burst of increased oxygen uptake. 4. The inhibition of respiration by Dio-9 is dependent upon the presence of inorganic phosphate or arsenate in the incubation medium. In the absence of phosphate, Dio-9 only stimulates the respiration which is further accelerated by dinitrophenol. 5. Dio-9 stimulates respiration in the presence of oligomycin or 2-n-heptyl-4hydroxyquinoline N-oxide in the absence of phosphate but not in its presence. 6. Arsenate-stimulated respiration is only slightly inhibited by Dio-9. However, the increased respiration produced by 2,4-dinitrophenol in the presence of arsenate is not obtained in the presence of Dio-9. 7. Dio-9 has a much smaller effect on sonicated mitochondria or on the Keilin and Hartree heart-muscle preparation. 8. Dio-9 causes a minor swelling of rat-liver mitochondria which is potentiated by inorganic phosphate. 9. The antibiotic has little effect on the 2,4-dinitrophenoMnduced ATPase of freshly prepared mitochondria although it inhibits the ATPase of aged mitochondria. IO. The possible mode of action of Dio-9 is discussed.
Biotechnology and applied biochemistry, 1994
A cell-free system prepared from polymorphonuclear neutrophils is capable of NADPH-dependent gene... more A cell-free system prepared from polymorphonuclear neutrophils is capable of NADPH-dependent generation of superoxide anion, but requires the simultaneous presence of plasma membranes, cytosol, arachidonate and guanosine 5'-[gamma-thio]triphosphate (GTP[S]). The isolated membranes from such a preparation are able to catalyse NADPH-dependent superoxide formation independently of added cytosol and activators. Such activated membranes, activated in the cell-free system, must consequently contain all of the essential components required by the oxidase for superoxide formation, including the NADPH-binding component. Arylazido-beta-alanyl-[32P]NADPH (3'-O-(3-[N-(4-azido-2-nitrophenyl)-amino] propionyl)-[32P]NADPH), an NADPH analogue and photoaffinity probe, is shown to act in the dark as a substrate for the oxidase activity in the activated membranes and an irreversible photodependent inhibitor following photoirradiation. The photoaffinity probe has been used to identify the speci...
The Journal of biological chemistry, Jan 10, 1971
The Biochemical journal, 1972
Characteristics of inorganic pyrophosphate synthesis from inorganic orthophosphate were examined ... more Characteristics of inorganic pyrophosphate synthesis from inorganic orthophosphate were examined in chromatophores of Rhodospirillum rubrum. The application of an ADP-glucose pyrophosphorylase-trapping system has shown in an unequivocal fashion that pyrophosphate is a product of a light-dependent reaction utilizing P(i) as the substrate. Only very limited pyrophosphate synthesis takes place in the dark. The rates of synthesis of both ATP and pyrophosphate were studied under conditions in which the membrane-bound adenosine triphosphatase and pyrophosphatase activities would normally make these substances unstable. The maximum rate of pyrophosphate synthesis was 25% of that for ATP synthesis, with maximum activation of pyrophosphate synthesis occurring at a lower light-intensity than that required for ATP synthesis. As a result, at low light-intensity the rate of pyrophosphate formation approached that of ATP. Maximal rates of synthesis of both pyrophosphate and ATP were attained only...
The Journal of biological chemistry, Jan 10, 1969
Biochimica et biophysica acta, Jan 24, 1965
1. Previous work indicating the specificity of hexylguanidines for oxidative phosphorylation at S... more 1. Previous work indicating the specificity of hexylguanidines for oxidative phosphorylation at Site I was confirmed. 50% inhibition was obtained with 0.02 μmole hexylguanidine per mg protein. ... 2. Synthalin (decamethylenediguanidine) completely inhibits the acceptor-...
The Biochemical journal, 1972
The preparation of ox heart myosin and its partial digestion with cellulose-bound papain is descr... more The preparation of ox heart myosin and its partial digestion with cellulose-bound papain is described. A procedure is outlined by which heavy meromyosin subfragment 1 can be covalently bound to a cellulose ion-exchange matrix. Attachment of heavy meromyosin subfragment 1 to the insoluble matrix results in a change in the ion specificity towards ATP hydrolysis. Unlike the soluble enzyme the bound form is activated by both Ca(2+) and Mg(2+). Maximal activation by Ca(2+) occurred at a lower concentration for the bound enzyme. Mg(2+) activates at a concentration which causes near-maximal inhibition of the Ca(2+)-activated adenosine triphosphatase (ATPase) of the non-bound enzyme. The Mg(2+)-activated ATPase of the bound enzyme was in turn inhibited by the presence of Ca(2+). The activation by Mg(2+) resembles the characteristic enzymic action of the actin-subfragment 1 complex.
Journal of supramolecular structure, 1975
The synthesis of ATP analogs containing a photoactive aryl azido grouping coupled to the 3' h... more The synthesis of ATP analogs containing a photoactive aryl azido grouping coupled to the 3' hydroxyl of ATP is described. The potential effectiveness of these analogs in the investigation of nucleotide-binding regions is outlined and this effectiveness demonstrated by their photodependent inhibition of subfragment 1 ATPase. The use of 14C-labeled azido ATP demonstrates an almost stoichiometric covalent binding of the analog. Because of their potential application to other systems, a number of reactions describing the reactivity of the 3' hydroxyl of the nucleotide ribose are outlined in an Appendix.
Journal of Protein Chemistry, 1986
Snake presynaptic toxins such as crotoxin, ß-bungarotoxin and taipoxin block neuromuscular transm... more Snake presynaptic toxins such as crotoxin, ß-bungarotoxin and taipoxin block neuromuscular transmission through inhibiting the release of acetylcholine by their phospholipase A 2 activities. On the other hand, many other phospholipase A 2 s show ...
Journal of biochemistry, molecular biology, and biophysics : JBMBB : the official journal of the Federation of Asian and Oceanian Biochemists and Molecular Biologists (FAOBMB), 2002
Functional reagents known to bring about the formation of a distinct membrane molecular complex o... more Functional reagents known to bring about the formation of a distinct membrane molecular complex of the subunits of cytochrome b(558) (gp 91(phox) and p22(phox)) were investigated for their influence on the O2- generating capability of liposome incorporated cytochrome b(558) preparations. One, ethyleneglycolbis[sulfo-succinimidylsuccinate], (sulfo-EGS) was found to inhibit O2- generation at concentrations which are known to result in cross-linking the two subunits of cytochrome b(558). Sulfosuccinimidyl [4-azidophenyldithio] propionate, (sulfo-SADP) on the other hand, was found to be a powerful inhibitor of the cytochrome b(558) dependent O2- production at concentrations not able to result in cross linking of the two subunits. Sulfo-SADP inhibits the cytochrome b(558) O2- production 50% at 25 microM, while sulfo-EGS requires 400 microM. For these reagents, the succinimidyl group of sulfo-SADP and sulfo-EGS is the reactive group, which inhibit irreversibly, cytochrome b(558) generatio...
Science, 1977
A photoaffinity labeling technique is described by which a tetrodotoxin analog is covalently boun... more A photoaffinity labeling technique is described by which a tetrodotoxin analog is covalently bound to receptor sites associated with the sodium pores of excitable membranes. The biological activity of the toxin analog is retained after the covalent binding reaction.
Medical Biochemistry, 2002
Journal of Protein Chemistry, 1986
Proceedings of the National Academy of Sciences, 1994
Polymorphonuclear neutrophil membranes, activated in a cell-free system, contain all of the essen... more Polymorphonuclear neutrophil membranes, activated in a cell-free system, contain all of the essential components required for superoxide formation including the NADPH-binding component. Arylazido-l-alanyl-[32P]NADPH-3'-0-{3-[N-(4-azido-2-nitrophenyl)aminoI propionyl}-[32P]NADPH-an NADPH analogue and photoaffinity probe, has been used to identify the specific NADPH binding component of the oxidase in activated membranes. A protein of about 52 kDa was photodependently labeled in the activated membranes by arylazldo-3-alanyl-[32P]NADPH. Specificity of labeling was indicated by the absence of such labeling in nonactivated membranes. The 52-kDa-labeled protein was the only isotopically labeled protein extracted from the labeled membranes with the chaotrope sodium perchlorate. Sodium perchlorate extraction of the 52-kDa protein from activated membranes correlates with the loss of the membranes' superoxide-generating capability. Reconstitution of the lost activity for sodium perchlorate-extracted membranes was accomplished by reincubating the extracted membranes with cytosol. It is proposed that the arylazido-,3-alanyl-[32PJNADPH-labeled protein of 52to 57-kDa present on the activated membranes is the NADPH-binding protein of the neutrophil superoxide-generating oxidase.
Biochem Biophys Res Commun, 1985
The purified mitochondrial NADH dehydrogenase enzyme has been shown to catalyze a rapid [4B-3H] N... more The purified mitochondrial NADH dehydrogenase enzyme has been shown to catalyze a rapid [4B-3H] NADH-H20 exchange reaction. When the enzyme is subjected to a single freeze-thaw cycle there is a complete loss of NADH dehydrogenation without a measurable decrease in the [4B-3H] NADH-H20 exchange. Complete loss of the [4B-3H] NADH-H20 exchange follows brief exposure to ultraviolet photoirradiation. The differential sensitivity of the water exchange reaction and the dehydrogenase activity suggests a direct involvement of the enzymes flavin cofactor in the catalysis of the [4B-3H] NADH-H20 exchange. Arylazido-S-alanyl NAD+ (A3'-0-{3-[N-4-azido-2-nitrophenyl)amino] propionyl}NAD+) is shown to be a potent photodependent inhibitor of the [4B-3H] NADH-H20 exchange activity following photoirradiation with visible light. This is consistent with the observed photodependent inhibition of the dehydrogenase activity by this photoprobe (Chen, S. and Guillory, R.J. (1981) J. Biol. Chem. 256, 8318-8323). The abbreviations used are: arylazido-S-alanyl NAD+, A3'-O-{3-[N-(4-azido-2nitrophenyl)amino]propionyl)NAD+, AcPyAD+, 3-acetyl pyridine adenine dinucleotide.
Proceedings of the National Academy of Sciences of the United States of America, 1994
Biochemistry Usa, 1997
The leukocyte iodonitrotetrazolium violet (INT) reductase activity of disrupted bovine polymorpho... more The leukocyte iodonitrotetrazolium violet (INT) reductase activity of disrupted bovine polymorphonuclear neutrophils is closely associated with the activation of the O 2-generating NADPH oxidase in a cell-free system. It is dependent upon NADPH, cytosolic factors, and amphiphiles (such as arachidonate), the same factors required for O 2-generation. Both O 2-generation and INT reductase activity are inhibited by phenylarsine oxide, an inhibitor of the activation of the NADPH oxidase [
Molecular Structure, Function, and Assembly of the ATP Synthases, 1989
Current Topics in Bioenergetics, 1979
Proceedings of the National Academy of Sciences of the United States of America, 1959
Biochimica et biophysica acta, Jan 26, 1964
I. The antibiotic Dio-9 inhibits the acceptor-controUed respiration of intact rat-liver mitochond... more I. The antibiotic Dio-9 inhibits the acceptor-controUed respiration of intact rat-liver mitochondria in a phosphate-containing medium. Essentially the same effects have been found with glutamate, fl-hydroxybutyrate, a-ketoglutarate, succinate and the ascorbate-tetramethyl-p-phenylenediamine system as substrate. 2. Inhibition of respiration by Dio-9 in a phosphate-containing system cannot be reversed by 2,4-dinitrophenol. However, if 2,4-dinitrophenol is added in uncoupling concentrations (o.I raM) prior to the antibiotic there is no inhibition of respiration by Dio-9. 3. The inhibition of respiration by Dio-9 is preceded by a short burst of increased oxygen uptake. 4. The inhibition of respiration by Dio-9 is dependent upon the presence of inorganic phosphate or arsenate in the incubation medium. In the absence of phosphate, Dio-9 only stimulates the respiration which is further accelerated by dinitrophenol. 5. Dio-9 stimulates respiration in the presence of oligomycin or 2-n-heptyl-4hydroxyquinoline N-oxide in the absence of phosphate but not in its presence. 6. Arsenate-stimulated respiration is only slightly inhibited by Dio-9. However, the increased respiration produced by 2,4-dinitrophenol in the presence of arsenate is not obtained in the presence of Dio-9. 7. Dio-9 has a much smaller effect on sonicated mitochondria or on the Keilin and Hartree heart-muscle preparation. 8. Dio-9 causes a minor swelling of rat-liver mitochondria which is potentiated by inorganic phosphate. 9. The antibiotic has little effect on the 2,4-dinitrophenoMnduced ATPase of freshly prepared mitochondria although it inhibits the ATPase of aged mitochondria. IO. The possible mode of action of Dio-9 is discussed.
Biotechnology and applied biochemistry, 1994
A cell-free system prepared from polymorphonuclear neutrophils is capable of NADPH-dependent gene... more A cell-free system prepared from polymorphonuclear neutrophils is capable of NADPH-dependent generation of superoxide anion, but requires the simultaneous presence of plasma membranes, cytosol, arachidonate and guanosine 5'-[gamma-thio]triphosphate (GTP[S]). The isolated membranes from such a preparation are able to catalyse NADPH-dependent superoxide formation independently of added cytosol and activators. Such activated membranes, activated in the cell-free system, must consequently contain all of the essential components required by the oxidase for superoxide formation, including the NADPH-binding component. Arylazido-beta-alanyl-[32P]NADPH (3'-O-(3-[N-(4-azido-2-nitrophenyl)-amino] propionyl)-[32P]NADPH), an NADPH analogue and photoaffinity probe, is shown to act in the dark as a substrate for the oxidase activity in the activated membranes and an irreversible photodependent inhibitor following photoirradiation. The photoaffinity probe has been used to identify the speci...
The Journal of biological chemistry, Jan 10, 1971
The Biochemical journal, 1972
Characteristics of inorganic pyrophosphate synthesis from inorganic orthophosphate were examined ... more Characteristics of inorganic pyrophosphate synthesis from inorganic orthophosphate were examined in chromatophores of Rhodospirillum rubrum. The application of an ADP-glucose pyrophosphorylase-trapping system has shown in an unequivocal fashion that pyrophosphate is a product of a light-dependent reaction utilizing P(i) as the substrate. Only very limited pyrophosphate synthesis takes place in the dark. The rates of synthesis of both ATP and pyrophosphate were studied under conditions in which the membrane-bound adenosine triphosphatase and pyrophosphatase activities would normally make these substances unstable. The maximum rate of pyrophosphate synthesis was 25% of that for ATP synthesis, with maximum activation of pyrophosphate synthesis occurring at a lower light-intensity than that required for ATP synthesis. As a result, at low light-intensity the rate of pyrophosphate formation approached that of ATP. Maximal rates of synthesis of both pyrophosphate and ATP were attained only...
The Journal of biological chemistry, Jan 10, 1969
Biochimica et biophysica acta, Jan 24, 1965
1. Previous work indicating the specificity of hexylguanidines for oxidative phosphorylation at S... more 1. Previous work indicating the specificity of hexylguanidines for oxidative phosphorylation at Site I was confirmed. 50% inhibition was obtained with 0.02 μmole hexylguanidine per mg protein. ... 2. Synthalin (decamethylenediguanidine) completely inhibits the acceptor-...
The Biochemical journal, 1972
The preparation of ox heart myosin and its partial digestion with cellulose-bound papain is descr... more The preparation of ox heart myosin and its partial digestion with cellulose-bound papain is described. A procedure is outlined by which heavy meromyosin subfragment 1 can be covalently bound to a cellulose ion-exchange matrix. Attachment of heavy meromyosin subfragment 1 to the insoluble matrix results in a change in the ion specificity towards ATP hydrolysis. Unlike the soluble enzyme the bound form is activated by both Ca(2+) and Mg(2+). Maximal activation by Ca(2+) occurred at a lower concentration for the bound enzyme. Mg(2+) activates at a concentration which causes near-maximal inhibition of the Ca(2+)-activated adenosine triphosphatase (ATPase) of the non-bound enzyme. The Mg(2+)-activated ATPase of the bound enzyme was in turn inhibited by the presence of Ca(2+). The activation by Mg(2+) resembles the characteristic enzymic action of the actin-subfragment 1 complex.
Journal of supramolecular structure, 1975
The synthesis of ATP analogs containing a photoactive aryl azido grouping coupled to the 3' h... more The synthesis of ATP analogs containing a photoactive aryl azido grouping coupled to the 3' hydroxyl of ATP is described. The potential effectiveness of these analogs in the investigation of nucleotide-binding regions is outlined and this effectiveness demonstrated by their photodependent inhibition of subfragment 1 ATPase. The use of 14C-labeled azido ATP demonstrates an almost stoichiometric covalent binding of the analog. Because of their potential application to other systems, a number of reactions describing the reactivity of the 3' hydroxyl of the nucleotide ribose are outlined in an Appendix.
Journal of Protein Chemistry, 1986
Snake presynaptic toxins such as crotoxin, ß-bungarotoxin and taipoxin block neuromuscular transm... more Snake presynaptic toxins such as crotoxin, ß-bungarotoxin and taipoxin block neuromuscular transmission through inhibiting the release of acetylcholine by their phospholipase A 2 activities. On the other hand, many other phospholipase A 2 s show ...
Journal of biochemistry, molecular biology, and biophysics : JBMBB : the official journal of the Federation of Asian and Oceanian Biochemists and Molecular Biologists (FAOBMB), 2002
Functional reagents known to bring about the formation of a distinct membrane molecular complex o... more Functional reagents known to bring about the formation of a distinct membrane molecular complex of the subunits of cytochrome b(558) (gp 91(phox) and p22(phox)) were investigated for their influence on the O2- generating capability of liposome incorporated cytochrome b(558) preparations. One, ethyleneglycolbis[sulfo-succinimidylsuccinate], (sulfo-EGS) was found to inhibit O2- generation at concentrations which are known to result in cross-linking the two subunits of cytochrome b(558). Sulfosuccinimidyl [4-azidophenyldithio] propionate, (sulfo-SADP) on the other hand, was found to be a powerful inhibitor of the cytochrome b(558) dependent O2- production at concentrations not able to result in cross linking of the two subunits. Sulfo-SADP inhibits the cytochrome b(558) O2- production 50% at 25 microM, while sulfo-EGS requires 400 microM. For these reagents, the succinimidyl group of sulfo-SADP and sulfo-EGS is the reactive group, which inhibit irreversibly, cytochrome b(558) generatio...
Science, 1977
A photoaffinity labeling technique is described by which a tetrodotoxin analog is covalently boun... more A photoaffinity labeling technique is described by which a tetrodotoxin analog is covalently bound to receptor sites associated with the sodium pores of excitable membranes. The biological activity of the toxin analog is retained after the covalent binding reaction.
Medical Biochemistry, 2002
Journal of Protein Chemistry, 1986
Proceedings of the National Academy of Sciences, 1994
Polymorphonuclear neutrophil membranes, activated in a cell-free system, contain all of the essen... more Polymorphonuclear neutrophil membranes, activated in a cell-free system, contain all of the essential components required for superoxide formation including the NADPH-binding component. Arylazido-l-alanyl-[32P]NADPH-3'-0-{3-[N-(4-azido-2-nitrophenyl)aminoI propionyl}-[32P]NADPH-an NADPH analogue and photoaffinity probe, has been used to identify the specific NADPH binding component of the oxidase in activated membranes. A protein of about 52 kDa was photodependently labeled in the activated membranes by arylazldo-3-alanyl-[32P]NADPH. Specificity of labeling was indicated by the absence of such labeling in nonactivated membranes. The 52-kDa-labeled protein was the only isotopically labeled protein extracted from the labeled membranes with the chaotrope sodium perchlorate. Sodium perchlorate extraction of the 52-kDa protein from activated membranes correlates with the loss of the membranes' superoxide-generating capability. Reconstitution of the lost activity for sodium perchlorate-extracted membranes was accomplished by reincubating the extracted membranes with cytosol. It is proposed that the arylazido-,3-alanyl-[32PJNADPH-labeled protein of 52to 57-kDa present on the activated membranes is the NADPH-binding protein of the neutrophil superoxide-generating oxidase.