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H Rochat

Publications and source records attributed to H Rochat.

At least 199 records · Page 11Linked to original sources

Covalent structure of the insect toxin of the North African scorpion Androctonus australis Hector.

The complete covalent structure of the insect toxin purified from the venom of the North-African scorpion Androctonus australis Hector was described. Its amino acid sequence was established by phenylisothiocyanate degradation of several protein derivatives and proteolytic fragments in a liquid protein sequencer using either a "protein" or a "peptide" program. The position of the four disulfide bridges were deduced by analysis of proteolytic peptides before and after diperformic oxidation, and by partial labeling of the half cystine residues with [14C]-iodoacetic acid and determining the specific radioactivities of the S-[14C]-carboxymethylated phenylthiohydantoin cysteines. The sequences of the insect and mammal toxins from scorpions can be aligned with homology with the positions of seven half-cystine residues as registers. The mammal and insect toxins have three disulfide bridges at homologous positions. The mammal and insect toxins have three disulfide bridges at homologous positions. The fourth bridge is different in that Cys12 in mammal toxin II is replaced by Cys38 in the insect toxin. It is likely that the position of the disulfide bridges is the same for all scorpion neurotoxins active on mammals. We believe that the shift of one half-cystine residue in the insect toxin may induce a conformational change in the structure of the protein, which, in turn, may partially account for the total specificity of this toxin for insect nervous system.

Amino Acid Sequence↗

The 1H nuclear-magnetic-resonance spectra and spatial structure of the Naja mossambica mossambica neurotoxin III.

The proton NMR spectra at 300 MHz of neurotoxin III from venom of Naja mossambica mossambica are reported. By the use of double resonance techniques, pH dependence chemical shifts, isotope labeling technique, and comparison with homologous neurotoxins all proton signals in the aromatic and methyl regions as well as epsilon-CH2 proton signals of some lysine residues have been assigned to individual amino acid residues and their spatial microenvironment has been determined. The results deduced on the solution structure of neurotoxin III are in complete agreement with the crystal structure of sea snake erabutoxins as well as with the previously established backbone folding and inter-residue interactions for the Naja naja oxiana short-chain neurotoxin in solution. In addition evidence has been obtained (a) that the conformation of the beta turn in the 31-34 segment depends on the ionization state of Asp-31 and His-32 side chain groups and (b) that an intricate electrostatic interaction exists in a system of ionogenic groups of the invariant Lys-27, Lys-47, Asp-31, Arg-33, Glu-38 and His-32 residues. These aspects of dynamic conformation are related to an interaction mechanism of a neurotoxin molecule and a nicotinic acetylcholine receptor.

Amino Acids↗

Penetration of phospholipid monolayers by cardiotoxins.

The monomolecular film technique was used to compare the specific interaction of four cardiotoxins from Naja mossambica mossambica with different phospholipids. We were able to demonstrate the interaction of cardiotoxins (10(-7) M) with both neutral and negatively charged phospholipids up to very high surface pressures (45 dyn/cm). In the presence of a phospholipid monolayer, the surface activity of cardiotoxins became much greater than that observed at the air-water interface. Neurotoxins of the same venom do not penetrate a phospholipid film, even at low surface pressure (15 dyn/cm). The apparent molecular area of cardiotoxin III during its insertion into a negatively charged phospholipid film was quantitatively defined. As a function of surface pressure of the membrane around 25 dyn/cm, cardiotoxins may exist in two different configurations, "flat" (1400 A2) or "edgewise" (420 A2). This result could account for the lytic activity of this type of toxin.

Cobra Cardiotoxin Proteins↗

Isolation and partial structural characterization of chicken pancreatic colipase.

Colipase has been isolated from acidic extracts of chicken pancreatic tissue homogenized with Triton X-100. The cofactor fully activates bile salt inhibited mammalian lipases. The amino terminal sequence of the avian protein has been determined up to position 39 and compared to the homologous region of the mammalian colipases (pig, horse, man) previously studied. From this comparison, it appears that a high degree of homology exists between the proteins.

Amino Acid Sequence↗

Structural studies on yeast 3-phosphoglycerate kinase. Linear arrangement of the CNBr fragments, partial amino acid sequence of the inner part of the polypeptide chain, and analyses of the N-terminal domain of the protein.

The purpose of this work was to contribute to the study of the covalent structure of yeast 3-phosphoglycerate kinase. First, we undertook the complete alignment of the four fragments produced by cyanogen-bromide cleavage and which constitute the intact protein; we then established the total amino acid sequence of a 30-residue peptide and the N-terminal sequence of a 65-residue peptide. Second, we analyzed the acetylated state of the protein. The analyses of the acid fraction "P" obtained after digestion of 3-phosphoglycerate kinase by pronase enabled us to determine the N-terminal sequence of this enzyme as N-acetylserylglycine. Third, we isolated, purified and analyzed seven tryptic peptides from a fragment containing 102 amino acids coming from the N-terminal end of the protein. The peptides occupying the N- and C-terminal ends of this fragment were also identified.

Amino Acid Sequence↗

Binding of scorpion neurotoxins to chick embryonic heart cells in culture and relationship to calcium uptake and membrane potential.

Stimulation of 45Ca uptake by scorpion neurotoxins in cultured chick embryonic heart cells has been shown to be directly linked to their effect on sodium channels. This property was used to compare the activity of 15 neurotoxins from five different species to their lethal effect in the mouse and immunological properties. As scorpion neurotoxins, the alkaloid neurotoxin veratridine enhanced 45Ca uptake, and an apparent positive cooperativity between the two drugs was observed. 125I-Labeled toxin II from the scorpion Androctonus australis Hector was shown to bind to chick heart cells specifically, saturably, and reversibly with high affinity (KD = 1--3 nM in sodium-free medium) and low capacity (10--20 fmol/mg cell protein). As shown by 45Ca uptake and radioactive toxin binding experiments, the affinity of scorpion neurotoxin to heart cell receptors was dependent on external K+ concentration. Toxin binding was lowered by increasing Na+ concentration in the medium and was abolished by veratridine in a sodium (140 mM) containing medium. As previously reported for neuroblastoma cells, all these results are in agreement with the membrane potential dependence of scorpion neurotoxin affinity for its membrane receptor.

Animals↗

Amino acid sequence of neurotoxin III of the scorpion Androctonus austrialis Hector.

The amino acid sequence of neurotoxin III, purified from the venom of the North African scorpion Androctonus australis Hector, has been determined by Edman degradation using a liquid-phase sequencer. Carboxypeptidase A hydrolyses confirmed not only the sequence of the five last residues but also the presence of a free alpha-carboxylic group at the C-terminus. Edman degradation was conducted on one hand with the Quadrol [N,N,N',N'-tetrakis(2-hydroxypropyl)ethylene diamine] program and S-alkylated protein before or after coupling with sulfophenylisothiocynate (the first 34 residues were thus identified), on the other hand on tryptic and chymotryptic peptides with a dimethylbenzylamine program (residues 1--23 and 31--34 were confirmed, the positions of residues 35-64 were established). Neurotoxin III was found to belong to the same group of scorpion toxins active on mammals as neurotoxin I purified from the same venom (50 homologous positions exist in the two proteins).

Amino Acid Sequence↗

Parvalbumins from coelacanth muscle. II. Amino acid sequence of the two less acidic components.

The primary structure of the two less acidic parvalbumins (pI = 5.44 and pI = 4.95) from coelacanth muscle (Latimeria chalumnae) has been determined. They differ only by the presence or absence of a N-terminal blocking group. By the use of the automatic degradation, 69 amino acids could be placed unambiguously in the N-terminal part and 24 amino acids following the single arginine 75. Tryptic peptides were used to establish the sequence and the position of the remaining residues. The two parvalbumins examined belong to the alpha-lineage, and the rate of their molecular evolution is comparable to that found in other vertebrates.

Amino Acid Sequence↗