References of "Ghuysen, Jean-Marie"
     in
Bookmark and Share    
Full Text
Peer Reviewed
See detailMode of Membrane Insertion and Sequence of a 32-Amino Acid Peptide Stretch of the Penicillin-Binding Protein 4 of Enterococcus Hirae
Jacques, Philippe; el Kharroubi, Aboubaker; Van Beeumen, Jozef et al

in FEMS Microbiology Letters (1991), 66(2), 119-123

Analysis of water-soluble derivatives of the Enterococcus hirae 75-kDa membrane-bound penicillin-binding protein 4 (PBP4) has yielded the amino acid sequence of a 32-amino acid polypeptide stretch. This ... [more ▼]

Analysis of water-soluble derivatives of the Enterococcus hirae 75-kDa membrane-bound penicillin-binding protein 4 (PBP4) has yielded the amino acid sequence of a 32-amino acid polypeptide stretch. This peptide is similar to peptide segments known to occur in the N-terminal domain of high-Mr PBPs of class B. The E. hirae PBP4 probably belongs to the same class. It is anchored in the membrane at the N-terminus of the polypeptide chain. [less ▲]

Detailed reference viewed: 16 (0 ULg)
Full Text
Peer Reviewed
See detailA standard numbering scheme for the class A beta-lactamases
Ambler, R. P.; Coulson, A. F. W.; Frère, Jean-Marie ULg et al

in Biochemical Journal (1991), 276(Pt 1), 269-270

Detailed reference viewed: 27 (2 ULg)
Full Text
Peer Reviewed
See detailRecherche fondamentale et médecine
Ghuysen, Jean-Marie ULg

in Revue Médicale de Liège (1991), 46(3), 115-122

Detailed reference viewed: 6 (0 ULg)
Full Text
Peer Reviewed
See detailPolarization correction to the electrostatic potential at the CNDO and the ab initio level. Influence of the basis set expansion
Dehareng, Dominique ULg; Dive, Georges ULg; Ghuysen, Jean-Marie ULg

in Theoretica Chimica Acta (1991), 79(2), 141-152

The influence of the basis set on the electrostatic potential corrected for polarization has been studied for H2S, CH3SH and COHCH2SH. The position and deepness of the minima and the height of the barrier ... [more ▼]

The influence of the basis set on the electrostatic potential corrected for polarization has been studied for H2S, CH3SH and COHCH2SH. The position and deepness of the minima and the height of the barrier between symmetric minima is discussed at both the deorthogonalized CNDO/2 and ab initio levels within STO-3G, 3-21G, 4-31G, 6-31G and 6-311G basis sets. The calculation of the electrostatic potential and corrected one using CNDO deorthogonalized coefficients including 3d orbitals has been applied at the first time on sulfur-containing molecules. The influence of polarization and diffuse functions has also been analysed and the incidence of the polarization correction on the relative proton affinity in NH2(CH2)3NHCH3 and in the adenine molecule has been investigated at the CNDO and ab initio levels. At both levels, the relative proton affinity of several basic sites in the same molecule can be qualitatively expressed without inclusion of the polarization correction except in the case of substituted amines. [less ▲]

Detailed reference viewed: 16 (2 ULg)
Full Text
Peer Reviewed
See detailConformational analysis of β and γ-lactam antibiotics
Lamotte, Josette ULg; Dive, Georges ULg; Ghuysen, Jean-Marie ULg

in European Journal of Medicinal Chemistry (1991), 26(1), 43-50

Geometry optimization, superimposition searches and conformational analysis carried on several lactam antibiotics show that reactivity with the active-site serine penicillin-binding proteins is related to ... [more ▼]

Geometry optimization, superimposition searches and conformational analysis carried on several lactam antibiotics show that reactivity with the active-site serine penicillin-binding proteins is related to a particular spatial disposition of 2 flanking functional groups - namely a C = O or C-OH on 1 side and a carboxylate on the other - with respect to the central scissile amide bond. Such a binding entity is found in one of the most stable conformers of the tripeptide diacetyl-L-Lys-D-Ala-D-Ala conferring substrate activity, and in benzylpenicillin, cephapyrin, thienamycin, gamma-lactam, the 6-spiro-epoxypenicillin S and in one epimer of lactivicin, conferring inactivating potency. This binding entity generates a particular electronic distribution and the fact that it is conserved in compounds belonging to very different chemical families strongly suggests that it is an important feature required for enzyme recognition. [less ▲]

Detailed reference viewed: 98 (6 ULg)
Full Text
Peer Reviewed
See detailSerine β-lactamases and penicillin-binding proteins
Ghuysen, Jean-Marie ULg

in Annual Review of Microbiology (1991), 45

Detailed reference viewed: 32 (0 ULg)
Full Text
Peer Reviewed
See detailAcyltransferase activities of the high-molecular-mass essential penicillin-binding proteins
Adam, Maggy; Damblon, Christian ULg; Jamin, Marc et al

in Biochemical Journal (1991), 279(Part 2), 601-604

The high-molecular-mass penicillin-binding proteins (HMM-PBPs), present in the cytoplasmic membranes of all eubacteria, are involved in important physiological events such as cell elongation, septation or ... [more ▼]

The high-molecular-mass penicillin-binding proteins (HMM-PBPs), present in the cytoplasmic membranes of all eubacteria, are involved in important physiological events such as cell elongation, septation or shape determination. Up to now it has, however, been very difficult or impossible to study the catalytic properties of the HMM-PBPs in vitro. With simple substrates, we could demonstrate that several of these proteins could catalyse the hydrolysis of some thioesters or the transfer of their acyl moiety on the amino group of a suitable acceptor nucleophile. Many of the acyl-donor substrates were hippuric acid or benzoyl-D-alanine derivatives, and their spectroscopic properties enabled a direct monitoring of the enzymic reaction. In their presence, the binding of radioactive penicillin to the PBPs was also inhibited. [less ▲]

Detailed reference viewed: 21 (3 ULg)
Full Text
Peer Reviewed
See detailCharacterization of an Enterococcus Hirae Penicillin-Binding Protein 3 with Low Penicillin Affinity
Piras, Graziella; El Kharroubi, Aboubaker; van Beeumen, Jozef et al

in Journal of Bacteriology (1990), 172(12), 6856-6862

Enterococcus hirae S185, a clinical isolate from swine intestine, exhibits a relatively high resistance to penicillin and contains two 77-kDa penicillin-binding proteins 3 of high (PBP 3s) and low (PBP 3r ... [more ▼]

Enterococcus hirae S185, a clinical isolate from swine intestine, exhibits a relatively high resistance to penicillin and contains two 77-kDa penicillin-binding proteins 3 of high (PBP 3s) and low (PBP 3r) affinity to penicillin, respectively. A laboratory mutant S185r has been obtained which overproduces PBP 3r and has a highly increased resistance to penicillin. Peptide fragments specifically produced by trypsin and SV8 protease digestions of PBP 3r were isolated, and the amino acid sequences of their amino terminal regions were determined. On the basis of these sequences, oligonucleotides were synthesized and used as primers to generate, by polymerization chain reaction, a 233-bp DNA fragment the sequence of which translated into a 73-amino-acid peptide segment of PBP 3r. These structural data led to the conclusion that the E. hirae PBP 3r and the methicillin-resistant staphylococcal PBP 2' are members of the same class of high-Mr PBPs. As shown by immunological tests, PBP 3r is not related to PBP 3s but, in contrast, is related to the 71-kDa PBP 5 of low penicillin affinity which is responsible for penicillin resistance in E. hirae ATCC 9790 and R40. [less ▲]

Detailed reference viewed: 13 (3 ULg)
Full Text
Peer Reviewed
See detailβ-Lactamase expression in Streptomyces cacaoi
Urabe, Hiroaki; Lenzini, Mauro V; Mukaide, Masakazu et al

in Journal of Bacteriology (1990), 172(11), 6427-6434

Plasmids were prepared by inserting genomic DNA fragments from Streptomyces cacaoi within the mel gene of plasmid pIJ702. The inserted DNA fragments contain the beta-lactamase-encoding bla gene and ... [more ▼]

Plasmids were prepared by inserting genomic DNA fragments from Streptomyces cacaoi within the mel gene of plasmid pIJ702. The inserted DNA fragments contain the beta-lactamase-encoding bla gene and upstream nucleotide sequences of various lengths. The transcription start point of bla was identified by nuclease S1 mapping. Upstream nucleotide sequences of sufficient lengths had an enhancing effect on beta-lactamase production by the Streptomyces host. The dot blot hybridization assay revealed that this effect was exerted at the transcriptional level. Experimental evidence strongly suggests that the underlying mechanism involves, at least in part, one or several trans-acting elements. In one of the constructs, in which the upstream nucleotide sequence was reduced to 0.3 kb, the bla promoter was present but the bla gene was expressed by readthrough from a promoter, possibly the mel promoter, of the pIJ702 vector. [less ▲]

Detailed reference viewed: 11 (3 ULg)
Full Text
Peer Reviewed
See detailEngineering a Novel β-Lactamase by a Single Point Mutation
Jacob, F.; Joris, Bernard ULg; Dideberg, O. et al

in Protein Engineering (1990), 4(1), 79-86

beta-Lactamases are widespread and efficient bacterial enzymes which play a major role in bacterial resistance to penicillins and cephalosporins. In order to elucidate the role of the residues lying in a ... [more ▼]

beta-Lactamases are widespread and efficient bacterial enzymes which play a major role in bacterial resistance to penicillins and cephalosporins. In order to elucidate the role of the residues lying in a conserved loop of the enzymatic cavity of the active-site serine Streptomyces albus G beta-lactamase, modified proteins were produced by oligo-directed mutagenesis. Mutation of Asn116, which lies on one side of the active site cavity pointing to the substrate-binding site, into a serine residue resulted in spectacular modifications of the specificity profile of the enzyme. That replacement yielded an enzyme with a nearly unchanged activity towards good penicillin substrates. In sharp contrast its efficiency in hydrolysing cephalosporins was drastically reduced, the best substrates suffering the largest decrease in the second-order rate constant for serine acylation. In fact that single mutation generated a truly new enzyme behaving exclusively as a penicillinase, a situation which is never encountered to the same degree in any of the numerous naturally occurring variants of class A beta-lactamases. [less ▲]

Detailed reference viewed: 12 (3 ULg)
Full Text
Peer Reviewed
See detailMembrane Topology, Structure, and Functions of the Penicillin-Interactive Proteins
Ghuysen, Jean-Marie ULg

in Biotechnology & Applied Biochemistry (1990, October), 12(5), 468-472

Detailed reference viewed: 14 (1 ULg)
Full Text
Peer Reviewed
See detailCloning, Nucleotide Sequence and Amplified Expression of the Gene Encoding the Extracellular Metallo (Zn) Dd-Peptidase of Streptomyces Albus G
Duez, Colette ULg; Lakaye, Bernard ULg; Houba, Simone et al

in FEMS Microbiology Letters (1990), 71(1-2), 215-219

The gene encoding the extracellular metallo (Zn) DD-peptidase of Streptomyces albus G has been cloned in Escherichia coli DH5 alpha MCR via pBR322 or 325, and then transferred into Streptomyces lividans ... [more ▼]

The gene encoding the extracellular metallo (Zn) DD-peptidase of Streptomyces albus G has been cloned in Escherichia coli DH5 alpha MCR via pBR322 or 325, and then transferred into Streptomyces lividans TK24 via pIJ486, with substantial amplification of the expressed DD-peptidase. The gene has the information for the synthesis of a 255 amino acid precursor, the amino terminal region of which has the characteristic features of a signal peptide. The primary structure as deduced from nucleotide sequencing confirms that previously determined by chemical methods except for the occurrence of an Asp instead of Asn at position 1 and an additional Ala immediately downstream of Pro67. [less ▲]

Detailed reference viewed: 8 (0 ULg)
Full Text
Peer Reviewed
See detailTranscriptional Analysis of the dd-Peptidase/Penicillin-Binding Protein-Encoding dac Gene of Streptomyces R61: Use of the Promoter and Signal Sequences in a Secretion Vector
Piron-Fraipont, Claudine; Lenzini, Mauro V; Dusart, Jean et al

in Molecular & General Genetics (1990), (223), 114-120

The promoter region of the gene encoding the extracellular DD-peptidase/penicillin-binding protein of Streptomyces R61 has been identified by in vivo promoter probing and S1 mapping. A secretion vector ... [more ▼]

The promoter region of the gene encoding the extracellular DD-peptidase/penicillin-binding protein of Streptomyces R61 has been identified by in vivo promoter probing and S1 mapping. A secretion vector, pDML116, was constructed by inserting into the multicopy Streptomyces plasmid pIJ702, a 247 bp DNA sequence that contained the transcriptional, translational and secretory signals and the 12 amino acid N-terminal region-encoding sequence of the mature Streptomyces DD-peptidase/penicillin-binding protein. Insertion, downstream of this 247 bp segment, of the Streptomyces R61 DD-peptidase-encoding gene or the Escherichia coli R-TEM beta-lactamase-encoding gene yielded plasmids pDML120 and pDML128, respectively, which allowed expression and secretion of the relevant enzymes by Streptomyces lividans. The maximal secretion levels obtained were 42 mg protein/ml for the autologous Streptomyces DD-peptidase and 0.9 mg protein/ml for the heterologous E. coli beta-lactamase. [less ▲]

Detailed reference viewed: 13 (3 ULg)
Full Text
Peer Reviewed
See detailElectrostatic potential maps at the quantum chemistry level of the active sites of the serine peptidases, α-chymotrypsin and subtilisin
Lamotte-Brasseur, Josette; Dive, Georges ULg; Dehareng, Dominique ULg et al

in Journal of Theoretical Biology (1990), 145(2), 183-98

The electronic properties of the active-sites of the structurally unrelated serine peptidases, alpha-chymotrypsin and subtilisin, have been expressed in the form of three-dimensional electrostatic ... [more ▼]

The electronic properties of the active-sites of the structurally unrelated serine peptidases, alpha-chymotrypsin and subtilisin, have been expressed in the form of three-dimensional electrostatic potential maps derived from integrals calculated at the quantum chemistry level. As a consequence of the asymmetrical distribution of the secondary structures that occur within a 7 A sphere around the serine of the catalytic triad, the active sites are highly polarized entities and exhibit large dipole moments. One part of the active sites generates a nucleophilic suction-pump. Its isocontour at -10 kcal mol-1 defines an impressive, negatively-charged volume which bears a narrow channel in the immediate vicinity of the active-site serine 195 in alpha-chymotrypsin or 221 in subtilisin. In native alpha-chymotrypsin, there is a perfect complementation between this nucleophilic suction-pump and the positively-charged electrophilic hole that is generated by the backbone NH of Ser 195 and Gly 193. In subtilisin, generation of the complementing electrophilic hole requires binding of a carbonyl donor ligand and may be achieved by rotation of the side-chain amide of Asn 155 towards the backbone NH of Ser 221. Small variations in the atomic co-ordinates of alpha-chymotrypsin used for the calculations, the presence of water molecules in its active site and the occurrence of point mutations in the amino acid sequence of subtilisin have little effects on the shape and characteristics of the electrostatic potential. [less ▲]

Detailed reference viewed: 27 (11 ULg)
Full Text
Peer Reviewed
See detailExpression in Escherichia Coli of the Carboxy Terminal Domain of the Blar Sensory-Transducer Protein of Bacillus Licheniformis as a Water-Soluble Mr 26,000 Penicillin-Binding Protein
Joris, Bernard ULg; Ledent, P.; Kobayashi, T. et al

in FEMS Microbiology Letters (1990), 58(1), 107-113

A cloning vector has been constructed which allows production and export by Escherichia coli of the Met346-Arg601 carboxy terminal domain of the 601 amino acid BLAR sensory-transducer involved in beta ... [more ▼]

A cloning vector has been constructed which allows production and export by Escherichia coli of the Met346-Arg601 carboxy terminal domain of the 601 amino acid BLAR sensory-transducer involved in beta-lactamase inducibility in Bacillus licheniformis. The polypeptide, referred to as BLAR-CTD, accumulates in the periplasm of E. coli in the form of a water-soluble, Mr 26,000 penicillin-binding protein. These data and homology searches suggest that BLAR has a membrane topology similar to that of other sensory-transducers involved in chemotaxis. [less ▲]

Detailed reference viewed: 23 (7 ULg)
Full Text
Peer Reviewed
See detailThe Life-Cycle Proteins Roda of Escherichia Coli and Spove of Bacillus Subtilis Have Very Similar Primary Structures
Joris, Bernard ULg; Dive, Georges ULg; Henriques, A. et al

in Molecular Microbiology (1990), 4(3), 513-517

Comparison of the predicted amino acid sequence of the cell-cycle RodA protein with the National Research Foundation protein sequence database shows that the 370-amino-acid RodA, a protein that is ... [more ▼]

Comparison of the predicted amino acid sequence of the cell-cycle RodA protein with the National Research Foundation protein sequence database shows that the 370-amino-acid RodA, a protein that is essential for wall elongation in Escherichia coli and maintenance of the rod shape of the cell, is highly analogous, in terms of primary structure, with the Bacillus subtilis SpoVE protein involved in stage V of sporulation. [less ▲]

Detailed reference viewed: 43 (2 ULg)
Full Text
Peer Reviewed
See detailIdentification of BlaR, the signal transducer for β-lactamase production in Bacillus licheniformis, as a penicillin-binding protein with strong homology to the OXA-2 β-lactamase (class D) of Salmonella typhimurium
Zhu, Ying-Fang; Curran, Ivan H. A.; Joris, Bernard ULg et al

in Journal of Bacteriology (1990), 172(2), 1137-1141

The blaR gene of Bacillus licheniformis encodes the signal transducer for induction of the class A beta-lactamase. The protein product, BlaR, has a hydrophilic carboxy region that binds beta-lactams and ... [more ▼]

The blaR gene of Bacillus licheniformis encodes the signal transducer for induction of the class A beta-lactamase. The protein product, BlaR, has a hydrophilic carboxy region that binds beta-lactams and shows high sequence homology to the class D beta-lactamases, particularly the OXA-2 beta-lactamase of Salmonella typhimurium. The BlaR-beta-lactam complex is stable and may provide the continuing stimulus needed for the prolonged production of the enzyme. [less ▲]

Detailed reference viewed: 22 (0 ULg)
Full Text
Peer Reviewed
See detailNucleotide sequence of the gene encoding the active-site serine β-lactamase from Actinomadura R39
Houba, Simone; Willem, Sabine; Duez, Colette ULg et al

in FEMS Microbiology Letters (1989), 65(3), 241-246

The gene encoding the extracellular, active-site serine beta-lactamase of Actinomadura R39, previously cloned into Streptomyces lividans, has the information for the synthesis of a 304 amino acid protein ... [more ▼]

The gene encoding the extracellular, active-site serine beta-lactamase of Actinomadura R39, previously cloned into Streptomyces lividans, has the information for the synthesis of a 304 amino acid protein, the amino terminal region of which has the characteristic features of a signal peptide. The Actinomadura R39 beta-lactamase is another member of the class A beta-lactamases. In particular, it shows high homology with the beta-lactamase of Bacillus licheniformis. [less ▲]

Detailed reference viewed: 14 (1 ULg)
Full Text
Peer Reviewed
See detailNucleotide sequences of the pbpX genes encoding the penicillin-binding proteins 2x from Streptococcus pneumoniae R6 and a cefotaxime-resistant mutant, C506
Laible, G.; Hakenbeck, R.; Sicard, M. A. et al

in Molecular Microbiology (1989), 3(10), 1337-1348

Development of penicillin resistance in Streptococcus pneumoniae is due to successive mutations in penicillin-binding proteins (PBPs) which reduce their affinity for beta-lactam antibiotics. PBP2x is one ... [more ▼]

Development of penicillin resistance in Streptococcus pneumoniae is due to successive mutations in penicillin-binding proteins (PBPs) which reduce their affinity for beta-lactam antibiotics. PBP2x is one of the high-Mr PBPs which appears to be altered both in resistant clinical isolates, and in cefotaxime-resistant laboratory mutants. In this study, we have sequenced a 2564 base-pair chromosomal fragment from the penicillin-sensitive S. pneumoniae strain R6, which contains the PBP2x gene. Within this fragment, a 2250 base-pair open reading frame was found which coded for a protein having an Mr of 82.35kD, a value which is in good agreement with the Mr of 80-85 kD measured by SDS-gel electrophoresis of the PBP2x protein itself. The N-terminal region resembled an unprocessed signal peptide and was followed by a hydrophobic sequence that may be responsible for membrane attachment of PBP2x. The corresponding nucleotide sequence of the PBP2x gene from C504, a cefotaxime-resistant laboratory mutant obtained after five selection steps, contained three nucleotide substitutions, causing three amino acid alterations within the beta-lactam binding domain of the PBP2x protein. Alterations affecting similar regions of Escherichia coli PBP3 and Neisseria gonorrhoeae PBP2 from beta-lactam-resistant strains are known. The penicillin-binding domain of PBP2x shows highest homology with these two PBPs and S. pneumoniae PBP2b. In contrast, the N-terminal extension of PBP2x has the highest homology with E. coli PBP2 and methicillin-resistant Staphylococcus aureus PBP2'. No significant homology was detected with PBP1a or PBP1b of Escherichia coli, or with the low-Mr PBPs. [less ▲]

Detailed reference viewed: 5 (0 ULg)
Full Text
Peer Reviewed
See detailActive-Site and Membrane Topology of the Dd-Peptidase/Penicillin-Binding Protein No. 6 of Enterococcus Hirae (Streptococcus Faecium) A.T.C.C. 9790
El Kharroubi, Aboubaker; Piras, Graziella; Jacques, Philippe et al

in Biochemical Journal (1989), 262(2), 457-462

The membrane-bound 43,000-Mr penicillin-binding protein no. 6 (PBP6) of Enterococcus hirae consists of a 30,000-Mr DD-peptidase/penicillin-binding domain and a approximately 130-residue C-terminal ... [more ▼]

The membrane-bound 43,000-Mr penicillin-binding protein no. 6 (PBP6) of Enterococcus hirae consists of a 30,000-Mr DD-peptidase/penicillin-binding domain and a approximately 130-residue C-terminal appendage. Removal of this appendage by trypsin proteolysis has no marked effect on the catalytic activity and penicillin-binding capacity of the PBP. Anchorage of the PBP in the membrane appears to be mediated by a short 15-20-residue stretch at the C-terminal end of the appendage. The sequence of the 50-residue N-terminal region of the PBP shows high degree of homology with the sequences of the corresponding regions of the PBPs5 of Escherichia coli and Bacillus subtilis. On this basis the active-site serine residue occurs at position 35 in the enterococcal PBP. [less ▲]

Detailed reference viewed: 8 (0 ULg)