A structure-activity study on the bradykinin B1 antagonist desArg10-HOE 140: The alanine scan

It has recently been shown that the biological activity of the second generation kinin B1 receptor selective antagonist, desArg10 HOE 140, can be improved by specific amino acid substitutions. Starting from this observation, we undertook a systematic structure-activity relationship study of this antagonist, based on the alanine-scan technique, in order to obtain useful information for the rational design of more analogues. Our data indicate that the sequence of desArg10 HOE 140 does not tolerate the replacement by Ala of most of its residues, with the exception of Ser in position 7 and, to a lesser extent, D-Arg in position 1 and Hyp in position 4. The most critical residues appear to be Pro in position 3 and the C-terminal dipeptide DTic-Oic; Ala replacement at these positions resultes in a total loss of activity. Moreover, replacement by Ala of Gly in position 5 reverts the activity of desArg10 HOE 140 to that of an agonist.     

Design,synthesis, and conformational studies of the hGM‐CSF derived peptide (13–27)–Gly–(75–87)

An analogue of the human granulocyte–macrophage colony‐stimulating factor (hGM‐CSF), hGM‐CSF(13–27)–Gly–(75–87) was synthesized by solid phase methodology. This analogue was designed to comprise helices A and C of the native growth factor, linked by a glycine bridge. Helices A and C form half of a four‐helix bundle motif in the crystal structure of the native factor and are involved in the interaction with α‐ and β‐chains of the heterodimeric receptor. A conformational analysis of the synthetic analogue by CD, two‐dimensional nmr spectroscopy, and molecular dynamics calculations is reported. The analogue is in a random structure in water and assumes a partially α‐helical conformation in a 1 : 1 trifluoroethanol/water mixture. The helix content in this medium is ∼ 70%. By 2D‐nmr spectroscopy, two helical segments were identified in the sequences corresponding to helices A and C. In addition to medium‐ and short‐range NOESY connectivities, a long‐range cross peak was found between the Cβ proton of Val¹⁶ and NH proton of His⁸⁷ (using the numbering of the native protein). Experimentally derived interproton distances were used as restraints in molecular dynamics calculations, utilizing the x‐ray coordinates as the initial structure. The final structure is characterized by two helical segments in close spatial proximity, connected by a loop region. This structure is similar to that of the corresponding domain in the x‐ray structure of the native growth factor in which helices A and C are oriented in an antiparallel fashion. The N‐terminal residues Gly–Pro of helix C are involved in an irregular turn connecting the two helical segments. As a consequence, helix C is appreciably shifted and slightly rotated with respect to helix A compared to the x‐ray structure of the native growth factor. These small differences in the topology of the two helices could explain the lower biological activity of this analogue with respect to that of the native growth factor. © 1999 John Wiley & Sons, Inc. Biopoly 50: 545–554, 1999     

Ulteriori indagini sul rapporto tra nichel e acidi malico e malonico in Alyssum

Abstract

Further contribution on the relationship between nickel and malic and malonic acids in Alyssum.—Further research has been carried out on extracts of leaves and seeds of A. bertolonii Desv., grown on serpentine and normal garden soil, in order to investigate the nature of the nickel compounds present in this plant. On the purified extracts, obtained with water and formic acid, malic and malonic acids have been determined through gas-chromatography of the trimethyl-sililderivatives; Ni, Ca, Mg and K have been analysed by atomic absorption spectrophotometry.

Both malic and malonic acids are present in the leaves of the plants collected on serpentine and therefore rich in nickel, whilst they are present only in small amounts in the plants grown in garden soil. Seeds contain mainly high levels of malic acid. Remarkable amounts of malic acid, related to a high nickel content, have been observed also in the leaves of another nickel accumulator in the genus Alyssum, i.e. A. serpyllifolium Desf. ssp. lusitanicum Dudley et P. Silva, endemic of Portugal serpentines.     

Influence of Mercury on the Amino Acidic Composition of Tobacco Leaves

In the course of the study on uptake of mercury by plants, tobacco was grown in the presence of mercury vapours and the amino acidic composition of the leaves was examined. The results showed that tobacco can take up more than 5,000 ppm of mercury through the leaves and store it, showing no symptom of poisoning. In relation to the absorption of mercury, an abnormal increase of cystine, glutamic acid and glycine was observed in the proteins of the leaves. A possible role of cystine in the metabolism of mercury is also discussed.     

On the Nature of Nickel Compounds in Alyssum Bertolonii Desv.-II

Fifteen derivatives of 5-substituted tetronic acid (VII) and five derivatives of 3-acetyl-5-substituted phenyltetramic acid (III) were prepared and their biological activities were investigated. Among the compounds tested, 3-carboethoxy derivatives of tetronic acid showed a remarkable stimulating effect on the growth of rice roots, while the compounds which were removed the carboethoxy group from their 3-position displayed a strong inhibitory effect on the growth of rice roots and stalks. The fungicidal activity against Asp. niger and the effects on pupation and emergence of the housefly were also investigated as to the compounds synthesized.     

Conformational studies on a synthetic C-terminal fragment of the alpha subunit of G(S) proteins

It has recently been reported that synthetic peptides corresponding to the C-terminal sequence of G alpha, can be used to study the molecular mechanisms of interaction between this protein and G protein coupled receptors (Hamm et al., Science, 1988, Vol. 241, pp. 832-835). A conformational analysis on a 11 amino acids peptide from the G alpha(S) C-terminus, G alpha(S)(384-394) (H-QRMHLRQYELL-OH), was performed by nmr spectroscopy and molecular modeling methods. Two-dimensional nmr spectra, recorded in hexafluoroacetone/water, a mixture with structure stabilizing properties, showed an unusually high number of nuclear Overhauser effects, forming significative pattern to the drawing of a secondary structure. Conformations consistent with experimental NOE distances were obtained through molecular dynamics and energy minimization methods. These calculations yielded two stable conformers corresponding to an alpha-turn and a type III beta-turn involving the last five C-terminal residues. Interestingly, the alpha-turn conformation was found to overlap with good agreement the crystallographic structure of the same fragment in the G alpha(S) protein.     

A structure-activity study on the bradykinin B1 antagonist desArg10-HOE 140: The alanine scan

Summary It has recently been shown that the biological activity of the second generation kinin B₁ receptor selective antagonist, desArg¹⁰ HOE 140, can be improved by specific amino acid substitutions. Starting from this observation, we undertook a systematic structure-activity relationship study of this antagonist, based on the alanine-scan technique, in order to obtain useful information for the rational design of more analogues. Our data indicate that the sequence of desArg¹⁰ HOE 140 does not tolerate the replacement by Ala of most of its residues, with the exception of Ser in position 7 and, to a lesser extent, D-Arg in position 1 and Hyp in position 4. The most critical residues appear to be Pro in position 3 and the C-terminal dipeptide Dtic-Oic; Ala replacement at these positions resultes in a total loss of activity. Moreover, replacement by Ala of Gly in position 5 reverts the activity of desArg¹⁰ HOE 140 to that of an agonist.