L. Debussche

Biosynthesis of vitamin B12: isolation of precorrin-6x, a metal-free precursor of the corrin macrocycle retaining five S-adenosylmethionine-derived peripheral methyl groups.

Thibaut, D, Debussche, L, Blanche, F

delta-Aminolevulinic acid and trimethylisobacteriochlorin are converted by cell-free protein preparations from Pseudomonas denitrificans into a metal-free pigment, precorrin-6x. This pigment, which...

Biosynthesis of vitamin B12: structure of precorrin-6x octamethyl ester.

Thibaut, D, Blanche, F, Debussche, L, Leeper, F J, Battersby, A R

13C-labeled precorrin-6x is biosynthesized by cell-free protein preparations from Pseudomonas denitrificans in separate experiments using delta-amino[5-13C]levulinic acid and the corresponding...

Purification of the two-enzyme system catalyzing the oxidation of the D-proline residue of pristinamycin IIB during the last step of pristinamycin IIA biosynthesis.

Thibaut, D, Ratet, N, Bisch, D, Faucher, D, Debussche, L, Blanche, F

High levels of conversion of 14C-labelled pristinamycin IIB (PIIB) to pristinamycin IIA (PIIA) were obtained in vivo in Streptomyces pristinaespiralis and in some other streptogramin A producers....

Purification of peptide synthetases involved in pristinamycin I biosynthesis.

Thibaut, D, Bisch, D, Ratet, N, Maton, L, Couder, M, Debussche, L, ...

Several assays of pristinamycin I synthetases based on adenylate or thioester formation were developed. Purification to near homogeneity of these enzymatic activities from cell extracts of...

Precorrin-6x reductase from Pseudomonas denitrificans: purification and characterization of the enzyme and identification of the structural gene.

Blanche, F, Thibaut, D, Famechon, A, Debussche, L, Cameron, B, Crouzet, J

Precorrin-6x reductase, which catalyzes the NADPH-dependent reduction of precorrin-6x to a dihydro derivative named precorrin-6y, was purified 14,300-fold to homogeneity with an 8% yield from...

The final step in the biosynthesis of hydrogenobyrinic acid is catalyzed by the cobH gene product with precorrin-8x as the substrate.

Thibaut, D, Couder, M, Famechon, A, Debussche, L, Cameron, B, Crouzet, J, ...

The final enzymatic reaction in the conversion of precorrin-6x to hydrogenobyrinic acid by cell-free protein preparations from Pseudomonas denitrificans was shown to be inhibited by hydrogenobyrinic...

Biosynthesis of vitamin B12 in Pseudomonas denitrificans: the biosynthetic sequence from precorrin-6y to precorrin-8x is catalyzed by the cobL gene product.

Blanche, F, Famechon, A, Thibaut, D, Debussche, L, Cameron, B, Crouzet, J

A protein catalyzing methylation at C-5 and C-15 and decarboxylation of the acetic acid side chain at C-12 on precorrin-6y to yield precorrin-8x was purified to homogeneity from a recombinant strain...

Biosynthesis of the corrin macrocycle of coenzyme B12 in Pseudomonas denitrificans.

Debussche, L, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Studies with cell-free protein preparations from a series of recombinant strains of Pseudomonas denitrificans demonstrated that precorrin-3 is converted into a further trimethylated intermediate,...

Assay, purification, and characterization of cobaltochelatase, a unique complex enzyme catalyzing cobalt insertion in hydrogenobyrinic acid a,c-diamide during coenzyme B12 biosynthesis in Pseudomonas denitrificans.

Debussche, L, Couder, M, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Hydrogenobyrinic acid a,c-diamide was shown to be the substrate of cobaltochelatase, an enzyme that catalyzes cobalt insertion in the corrin ring during the biosynthesis of coenzyme B12 in...

Purification and characterization of Cob(II)yrinic acid a,c-diamide reductase from Pseudomonas denitrificans.

Blanche, F, Maton, L, Debussche, L, Thibaut, D

An NADH-dependent flavoenzyme exhibiting cob(II)yrinic acid a,c-diamide reductase activity was purified 6,300-fold to homogeneity from Pseudomonas denitrificans and sequenced at its N terminus. This...

Biosynthesis of vitamin B12: stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide is catalyzed by one enzyme in Pseudomonas denitrificans.

Blanche, F, Couder, M, Debussche, L, Thibaut, D, Cameron, B, Crouzet, J

The cobalamin biosynthetic pathway enzyme that catalyzes amidation of 5'-deoxy-5'-adenosyl-cobyrinic acid a,c-diamide was purified to homogeneity from extracts of a recombinant strain of Pseudomonas...

A bifunctional protein from Pseudomonas denitrificans carries cobinamide kinase and cobinamide phosphate guanylyltransferase activities.

Blanche, F, Debussche, L, Famechon, A, Thibaut, D, Cameron, B, Crouzet, J

The two consecutive activities of the cobalamin biosynthetic pathway that catalyze the conversion of cobinamide to cobinamide phosphate (cobinamide kinase) and of cobinamide phosphate to...

Genetic analysis, nucleotide sequence, and products of two Pseudomonas denitrificans cob genes encoding nicotinate-nucleotide: dimethylbenzimidazole phosphoribosyltransferase and cobalamin (5'-phosphate) synthase.

Cameron, B, Blanche, F, Rouyez, M C, Bisch, D, Famechon, A, Couder, M, ...

Tn5 Sp(r) transposons have been inserted into the 8-kb Pseudomonas denitrificans DNA fragment from complementation group D, which carries cob genes. Genetic analysis and the nucleotide sequence...

Purification and partial characterization of Cob(I)alamin adenosyltransferase from Pseudomonas denitrificans.

Debussche, L, Couder, M, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Cob(I)alamin adenosyltransferase (EC 2.5.1.17) was purified to homogeneity from extracts of a Pseudomonas denitrificans recombinant strain and sequenced at its N terminus. It is a homodimer (each...

Purification and characterization of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Pseudomonas denitrificans.

Blanche, F, Debussche, L, Thibaut, D, Crouzet, J, Cameron, B

S-Adenosyl-L-methionine:uroporphyrinogen III methyltransferase (SUMT), the enzyme of the cobalamin biosynthetic pathway which catalyzes C methylation of uroporphyrinogen III, was purified about...

A Ras-GTPase-activating protein SH3-domain-binding protein.

Parker, F, Maurier, F, Delumeau, I, Duchesne, M, Faucher, D, Debussche, L, ...

We report the purification of a Ras-GTPase-activating protein (GAP)-binding protein, G3BP, a ubiquitously expressed cytosolic 68-kDa protein that coimmunoprecipitates with GAP. G3BP physically...

Antibodies to synthetic peptide from the residue 33 to 42 domain of c-Ha-ras p21 block reconstitution of the protein with different effectors.

Rey, I, Soubigou, P, Debussche, L, David, C, Morgat, A, Bost, P E, ...

Residues 32 to 40, which are conserved among ras proteins from different species, are likely to participate in interactions with the p21 effector system. With the goal of understanding the structural...

CTS1: a p53-derived chimeric tumor suppressor gene with enhanced in vitro apoptotic properties.

Conseiller, E, Debussche, L, Landais, D, Venot, C, Maratrat, M, Sierra, V, ...

The clinical potential of the p53 tumor suppressor gene is being evaluated currently for gene therapy of cancer. We have built a variant of wild-type p53, chimeric tumor suppressor 1 (CTS1), in which...

Purification and characterization of cobyrinic acid a,c-diamide synthase from Pseudomonas denitrificans.

Debussche, L, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Cobyrinic acid a,c-diamide synthase, which catalyzes the conversion of cobyrinic acid to cobyrinic acid a,c-diamide via the intermediate formation of cobyrinic acid c-monoamide, was purified 155-fold...

Assay and purification of S-adenosyl-L-methionine:precorrin-2 methyltransferase from Pseudomonas denitrificans.

Thibaut, D, Couder, M, Crouzet, J, Debussche, L, Cameron, B, Blanche, F

S-Adenosyl-L-methionine:precorrin-2 methyltransferase (SP2MT), which catalyzes the C-20 methylation of precorrin-2 to precorrin-3, was purified to homogeneity from extracts of a recombinant strain of...

Nucleotide sequence of a Pseudomonas denitrificans 5.4-kilobase DNA fragment containing five cob genes and identification of structural genes encoding S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase and cobyrinic acid a,c-diamide synthase.

Crouzet, J, Cauchois, L, Blanche, F, Debussche, L, Thibaut, D, Rouyez, M C, ...

A 5.4-kilobase DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment carries five different cob genes...

Genetic and sequence analysis of an 8.7-kilobase Pseudomonas denitrificans fragment carrying eight genes involved in transformation of precorrin-2 to cobyrinic acid.

Crouzet, J, Cameron, B, Cauchois, L, Rigault, S, Rouyez, M C, Blanche, F, ...

A 8.7-kilobase DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and the genetic analysis revealed that this fragment carries eight different cob...

The requirement for the p53 proline-rich functional domain for mediation of apoptosis is correlated with specific PIG3 gene transactivation and with transcriptional repression.

Venot, C, Maratrat, M, Dureuil, C, Conseiller, E, Bracco, L, Debussche, L

Wild-type p53 is a tumor suppressor gene which can activate or repress transcription, as well as induce apoptosis. The human p53 proline-rich domain localized between amino acids 64 and 92 has been...

Biosynthesis of vitamin B12: isolation of precorrin-6x, a metal-free precursor of the corrin macrocycle retaining five S-adenosylmethionine-derived peripheral methyl groups.

Thibaut, D, Debussche, L, Blanche, F

delta-Aminolevulinic acid and trimethylisobacteriochlorin are converted by cell-free protein preparations from Pseudomonas denitrificans into a metal-free pigment, precorrin-6x. This pigment, which...

Biosynthesis of vitamin B12: structure of precorrin-6x octamethyl ester.

Thibaut, D, Blanche, F, Debussche, L, Leeper, F J, Battersby, A R

13C-labeled precorrin-6x is biosynthesized by cell-free protein preparations from Pseudomonas denitrificans in separate experiments using delta-amino[5-13C]levulinic acid and the corresponding...

Purification of the two-enzyme system catalyzing the oxidation of the D-proline residue of pristinamycin IIB during the last step of pristinamycin IIA biosynthesis.

Thibaut, D, Ratet, N, Bisch, D, Faucher, D, Debussche, L, Blanche, F

High levels of conversion of 14C-labelled pristinamycin IIB (PIIB) to pristinamycin IIA (PIIA) were obtained in vivo in Streptomyces pristinaespiralis and in some other streptogramin A producers....

Purification of peptide synthetases involved in pristinamycin I biosynthesis.

Thibaut, D, Bisch, D, Ratet, N, Maton, L, Couder, M, Debussche, L, ...

Several assays of pristinamycin I synthetases based on adenylate or thioester formation were developed. Purification to near homogeneity of these enzymatic activities from cell extracts of...

Precorrin-6x reductase from Pseudomonas denitrificans: purification and characterization of the enzyme and identification of the structural gene.

Blanche, F, Thibaut, D, Famechon, A, Debussche, L, Cameron, B, Crouzet, J

Precorrin-6x reductase, which catalyzes the NADPH-dependent reduction of precorrin-6x to a dihydro derivative named precorrin-6y, was purified 14,300-fold to homogeneity with an 8% yield from...

The final step in the biosynthesis of hydrogenobyrinic acid is catalyzed by the cobH gene product with precorrin-8x as the substrate.

Thibaut, D, Couder, M, Famechon, A, Debussche, L, Cameron, B, Crouzet, J, ...

The final enzymatic reaction in the conversion of precorrin-6x to hydrogenobyrinic acid by cell-free protein preparations from Pseudomonas denitrificans was shown to be inhibited by hydrogenobyrinic...

Biosynthesis of vitamin B12 in Pseudomonas denitrificans: the biosynthetic sequence from precorrin-6y to precorrin-8x is catalyzed by the cobL gene product.

Blanche, F, Famechon, A, Thibaut, D, Debussche, L, Cameron, B, Crouzet, J

A protein catalyzing methylation at C-5 and C-15 and decarboxylation of the acetic acid side chain at C-12 on precorrin-6y to yield precorrin-8x was purified to homogeneity from a recombinant strain...

Biosynthesis of the corrin macrocycle of coenzyme B12 in Pseudomonas denitrificans.

Debussche, L, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Studies with cell-free protein preparations from a series of recombinant strains of Pseudomonas denitrificans demonstrated that precorrin-3 is converted into a further trimethylated intermediate,...

Assay, purification, and characterization of cobaltochelatase, a unique complex enzyme catalyzing cobalt insertion in hydrogenobyrinic acid a,c-diamide during coenzyme B12 biosynthesis in Pseudomonas denitrificans.

Debussche, L, Couder, M, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Hydrogenobyrinic acid a,c-diamide was shown to be the substrate of cobaltochelatase, an enzyme that catalyzes cobalt insertion in the corrin ring during the biosynthesis of coenzyme B12 in...

Purification and characterization of Cob(II)yrinic acid a,c-diamide reductase from Pseudomonas denitrificans.

Blanche, F, Maton, L, Debussche, L, Thibaut, D

An NADH-dependent flavoenzyme exhibiting cob(II)yrinic acid a,c-diamide reductase activity was purified 6,300-fold to homogeneity from Pseudomonas denitrificans and sequenced at its N terminus. This...

Biosynthesis of vitamin B12: stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide is catalyzed by one enzyme in Pseudomonas denitrificans.

Blanche, F, Couder, M, Debussche, L, Thibaut, D, Cameron, B, Crouzet, J

The cobalamin biosynthetic pathway enzyme that catalyzes amidation of 5'-deoxy-5'-adenosyl-cobyrinic acid a,c-diamide was purified to homogeneity from extracts of a recombinant strain of Pseudomonas...

A bifunctional protein from Pseudomonas denitrificans carries cobinamide kinase and cobinamide phosphate guanylyltransferase activities.

Blanche, F, Debussche, L, Famechon, A, Thibaut, D, Cameron, B, Crouzet, J

The two consecutive activities of the cobalamin biosynthetic pathway that catalyze the conversion of cobinamide to cobinamide phosphate (cobinamide kinase) and of cobinamide phosphate to...

Genetic analysis, nucleotide sequence, and products of two Pseudomonas denitrificans cob genes encoding nicotinate-nucleotide: dimethylbenzimidazole phosphoribosyltransferase and cobalamin (5'-phosphate) synthase.

Cameron, B, Blanche, F, Rouyez, M C, Bisch, D, Famechon, A, Couder, M, ...

Tn5 Sp(r) transposons have been inserted into the 8-kb Pseudomonas denitrificans DNA fragment from complementation group D, which carries cob genes. Genetic analysis and the nucleotide sequence...

Purification and partial characterization of Cob(I)alamin adenosyltransferase from Pseudomonas denitrificans.

Debussche, L, Couder, M, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Cob(I)alamin adenosyltransferase (EC 2.5.1.17) was purified to homogeneity from extracts of a Pseudomonas denitrificans recombinant strain and sequenced at its N terminus. It is a homodimer (each...

Purification and characterization of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Pseudomonas denitrificans.

Blanche, F, Debussche, L, Thibaut, D, Crouzet, J, Cameron, B

S-Adenosyl-L-methionine:uroporphyrinogen III methyltransferase (SUMT), the enzyme of the cobalamin biosynthetic pathway which catalyzes C methylation of uroporphyrinogen III, was purified about...

A Ras-GTPase-activating protein SH3-domain-binding protein.

Parker, F, Maurier, F, Delumeau, I, Duchesne, M, Faucher, D, Debussche, L, ...

We report the purification of a Ras-GTPase-activating protein (GAP)-binding protein, G3BP, a ubiquitously expressed cytosolic 68-kDa protein that coimmunoprecipitates with GAP. G3BP physically...

Antibodies to synthetic peptide from the residue 33 to 42 domain of c-Ha-ras p21 block reconstitution of the protein with different effectors.

Rey, I, Soubigou, P, Debussche, L, David, C, Morgat, A, Bost, P E, ...

Residues 32 to 40, which are conserved among ras proteins from different species, are likely to participate in interactions with the p21 effector system. With the goal of understanding the structural...

CTS1: a p53-derived chimeric tumor suppressor gene with enhanced in vitro apoptotic properties.

Conseiller, E, Debussche, L, Landais, D, Venot, C, Maratrat, M, Sierra, V, ...

The clinical potential of the p53 tumor suppressor gene is being evaluated currently for gene therapy of cancer. We have built a variant of wild-type p53, chimeric tumor suppressor 1 (CTS1), in which...

Purification and characterization of cobyrinic acid a,c-diamide synthase from Pseudomonas denitrificans.

Debussche, L, Thibaut, D, Cameron, B, Crouzet, J, Blanche, F

Cobyrinic acid a,c-diamide synthase, which catalyzes the conversion of cobyrinic acid to cobyrinic acid a,c-diamide via the intermediate formation of cobyrinic acid c-monoamide, was purified 155-fold...

Assay and purification of S-adenosyl-L-methionine:precorrin-2 methyltransferase from Pseudomonas denitrificans.

Thibaut, D, Couder, M, Crouzet, J, Debussche, L, Cameron, B, Blanche, F

S-Adenosyl-L-methionine:precorrin-2 methyltransferase (SP2MT), which catalyzes the C-20 methylation of precorrin-2 to precorrin-3, was purified to homogeneity from extracts of a recombinant strain of...

Nucleotide sequence of a Pseudomonas denitrificans 5.4-kilobase DNA fragment containing five cob genes and identification of structural genes encoding S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase and cobyrinic acid a,c-diamide synthase.

Crouzet, J, Cauchois, L, Blanche, F, Debussche, L, Thibaut, D, Rouyez, M C, ...

A 5.4-kilobase DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment carries five different cob genes...

Genetic and sequence analysis of an 8.7-kilobase Pseudomonas denitrificans fragment carrying eight genes involved in transformation of precorrin-2 to cobyrinic acid.

Crouzet, J, Cameron, B, Cauchois, L, Rigault, S, Rouyez, M C, Blanche, F, ...

A 8.7-kilobase DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and the genetic analysis revealed that this fragment carries eight different cob...

The requirement for the p53 proline-rich functional domain for mediation of apoptosis is correlated with specific PIG3 gene transactivation and with transcriptional repression.

Venot, C, Maratrat, M, Dureuil, C, Conseiller, E, Bracco, L, Debussche, L

Wild-type p53 is a tumor suppressor gene which can activate or repress transcription, as well as induce apoptosis. The human p53 proline-rich domain localized between amino acids 64 and 92 has been...