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J Dirkx

Publications and source records attributed to J Dirkx.

At least 19 recordsLinked to original sources
PubMed1979

Conformational studies on somatostatin. II. The C-terminal hexapeptide fragment.

The results of a conformational study on the C terminal hexapeptide of Somatostatin are presented. Semi-empirical energy calculations and high resolution NMR methods have been used to obtain information on the conformational properties of SRIF9-14 in [2H6]dimethylsulfoxide and 2H2O. It is concluded from the energy calculations that the peptide has an averaged conformation in which semi extended and folded structures are important. Only some of the folded conformations can explain the chemical shift differences between the amino acid residues Thr10 and Thr12 as a ring current shift by the Phe11 aromatic ring on Thr10. The nonequivalence is more pronounced in dimethyl-sulfoxide (0.23--0.15 ppm) where it decreases with increasing temperature towards the temperature independent value in 2H2O (0.03 ppm). This suggests that the folded conformations are somewhat predominant in dimethylsulfoxide solutions. In 2H2O the semi extended and folded structures are statistically equally important and the peptide is more flexible. A comparison with a study on the smaller fragments SRIF10-12 and SRIF10-13 which have similar conformational properties, demonstrates the usefulness of the fragment approach in conformational studies of peptides.

Amino Acid Sequence↗
PubMed1979

The conformational properties of some fragments of the peptide hormone somatostatin.

The conformational properties of some somatostatin fragments have been studied using high resolution NMR and semi-empirical calculations. The fragments are Thr10-Phe11, Phe11-Thr12, Thr10-Phe11-Thr12 and Thr10-Phe11-Thr12-Ser13. The results of high resolution 1H and 13C NMR using dimethylsulfoxide and 2H2O as solvents, combined with a new method for determining dihedral angles phi and psi from 13C and 1H spin lattice relaxation times are presented. A marked inequivalence of the two Thr10,12 residues is attributed to a shielding of the the Thr10 side chain by the Phe11 aromatic ring. The calculations show the existence of extended and folded low energy conformations in the tri and tetrapeptide. Only the folded structures give the observed shielding of Thr10. A temperature study of the tri and the tetrapeptide indicates that the folded structures are energetically the most favorable conformations at room temperature in dimethylsulfoxide. Increasing temperature reduces the nonequivalence of the Thr residues towards the differences that are observed in 2H2O. A detailed comparison of 3JalphaNH coupling constants and relaxation time measurements with the calculated conformations gives in general good agreement between both approaches. It is concluded that in these linear peptides, although several quite different low energy conformations exist, some of them are predominant. The continuity of both NMR parameters and calculated low energy conformations, when going from the smaller to the larger peptides, demonstrates the existence of structural properties far from the "random conformation".

Dimethyl Sulfoxide↗
PubMed1979

A program for generation and selection of possible conformations of cyclic molecules.

An algorithm has been developped for the computer generation of the possible conformations of non-planar cyclic molecules. This algorithm has been used successfully to select the known conformations of the ribose ring, of substituted cyclohexane and of disulfide bridged peptides. Its use may be extended to open molecules when the end-to-end distance is known or imposed.

Computers↗
PubMed1978

Isolation and characterization of the cyanogen bromide peptides from the B fragment of diphtheria toxin.

Homogeneous fragment B, obtained through nicking of diphtheria toxin with insoluble trypsin, was cleaved with cyanogen bromide in 70% formic acid. After citraconylation, the cleavage products were separated by gel filtration on Sephadex G--75 and purified by gel filtration, ion-exchange and thin-layer or paper chromatography. Six CNBr peptides were characterized, the composition of which account for the total amino acid content of fragment B. Their apparent molecular weights are: CB 1, 12 000; CB 2, 14 000; CB 3, 8000; CB 4a, 2400; CB 4b, 2200; CB 5, 2200. CB 4a is the NH2--terminal peptide; it contains the cysteine residue of the disulfide bridge linking fragment B to fragment A. CB 3 is the COOH--terminal peptide; it bears the disulfide bridge of fragment B. Characterization of two CNBr--derived overlapping peptides provided the positioning of CB 4b and CB 2 and allowed an alignment of the CNBr peptides of fragment B to be proposed.

Amino Acid Sequence↗
PubMed1977

Occurrence of tryptophan in the enzymically active site of diphtheria toxin fragment A.

Tryptophan residues of diphtheria toxin fragment A have been modified by 2-hydroxy-5-nitrobenzyl bromide. Fragment A loses its ability to inactivate ADP-ribosylation of the elongation factor 2, as a function of the number of residues modified. Modification of one tryptophan residue provides a dramatic loss of enzymic activity suggesting the presence of one essential residue of this type in fragment A. Examination of fingerprint maps of chymotryptic peptides of fragment A after reaction with 2-hydroxy-5-nitrobenzyl bromide allows us to identify this tryptophan residue at position 153 of the amino acid sequence. ORD and CD experiments joined to immunochemical studies seem to exclude the possibility that the observed decrease in the enzymic activity may be attributed to a conformational change. Finally, results of nicotinamide-adenine dinucleotide-binding measurements suggest that tryptophan 153 would be concerned with the elongation factor 2 binding site or with the catalytic site itself.

Benzyl Compounds↗