Determination of disulfide bridges in natural and recombinant insect defensin A

Lepage, Pierre; Bitsch, Francis; Roecklin, Dominique; Keppi, Elisabeth; Dimarcq, Jean‐Luc; Reichhart, Jean‐Marc; Hoffmann, Jules A.; Roitsch, C.; Dorsselaer, Alain Van

doi:10.1111/j.1432-1033.1991.tb15872.x

Cited by 28 publications

(22 citation statements)

References 16 publications

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“…Interestingly, moricin has no modified amino acid residues such as the ␣-amidation of C termini in cecropins, the hydroxylation of Lys residues in B. mori cecropins, the O-glycosylation of Thr residues in proline-rich antibacterial peptides (Bulet et al, 1993;Cociancich et al, 1994b;Hara and Yamakawa, 1995), or formation of intramolecular disulfide bonds in defensins (Kuzuhara et al, 1990;Lepage et al, 1990). These unique properties of moricin provide a favorable condition for the production of the peptide by chemical synthesis or by biotechnology using suitable protein expression vectors.…”

Section: Table III Effect Of the Novel Peptide On Viability Of S Aurmentioning

confidence: 99%

Moricin, a Novel Type of Antibacterial Peptide Isolated from the Silkworm, Bombyx mori

Hara

Yamakawa

1995

Journal of Biological Chemistry

163

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A novel antibacterial peptide that shows antibacterial activity against Staphylococcus aureus was isolated from the hemolymph of the silkworm, Bombyx mori. The novel peptide consisted of 42 amino acids and was highly basic. This peptide indicated no significant similarity with other antibacterial peptides. The peptide showed antibacterial activity against several Gram-negative and -positive bacteria and had a higher activity against Gram-positive bacteria than cecropin B 1 , a major antibacterial peptide of B. mori. The novel peptide was inducible by bacterial injection. These results suggest that the peptide is responsible for the antibacterial activity in B. mori against Gram-positive bacteria. The effects of the peptide on bacterial and liposomal membranes showed that a target of the peptide is the bacterial cytoplasmic membrane. The results also suggest that the N-terminal portion of the peptide, containing a predicted ␣-helix, is responsible for an increase in the membrane permeability. We propose the name "moricin" for this novel antibacterial peptide isolated from B. mori.

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Section: Table III Effect Of the Novel Peptide On Viability Of S Aurmentioning

confidence: 99%

Moricin, a Novel Type of Antibacterial Peptide Isolated from the Silkworm, Bombyx mori

Hara

Yamakawa

1995

Journal of Biological Chemistry

163

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“…This mass spectrum also contains two large peaks at mlz 565.0 and at mlz 1678.4 that we interpreted as fragments obtained from a tryptic peptide after rupture of an interchain disulfide bond. It has been shown that peptides consisting of two chains linked by a disulfide bridge undergo, in LSIMS, an intense fragmentation at the disulfide bridge which generates an [M+H]+ ion for each peptide chain [16]. (----) show all tryptic identified peptides ; the fragments marked with a full line ( -) show peptides identified after endoproteinase Glu-C cleavage.…”

Section: Endoproteinase Glu-cmentioning

confidence: 99%

“…From the known sequence of the CHH, one can predict which peptide could be generated by endoproteinase Glu-C digestion and only one solution was possible to explain the spectrum of the peptide from peak 6 [16]. One chain would be SCLKD (peptide 51-55; expected mass 564.2 Da) and the other DCYNLYRKPHVAAE (peptide 25 -38 ; expected mass 1677.8 Da).…”

Section: Endoproteinase Glu-cmentioning

confidence: 99%

Isolation and molecular characterization of a hyperglycemic neuropeptide from the sinus gland of the terrestrial isopod Armadillidium vulgare (Crustacea)

Martin

Sorokine

Dorsselaer

1993

European Journal of Biochemistry

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The major peptide from the sinus gland of the terrestrial isopod Armadillidium vulgare (Crustacea) has been extracted and purified by reverse-phase HPLC. This neuropeptide exhibited a high hyperglycemic activity and was therefore named A. vulgare crustacean hyperglycemic hormone (Am-CHH). Its average molecular mass measured by mass spectrometry was 8729.3 Da. Its complete amino acid sequence was determined by a combination of Edman degradation and mass spectrometry. The N-terminal amino acid was found to be unblocked, the C-terminal residue was found amidated and none of the other 72 residues was affected by any post-translational modification. Disulfide bond assignment was made unambigously by mass spectrometry and Edman degradation was performed on peptides produced by enzymatic cleavage. Relationships with other, similar neuropeptides from decapod sinus glands are discussed.The X-orgadsinus gland neurosecretory system located in the protocerebrum is the major neuroendocrine center in the isopod Armadillidium vulgare [l]. The sinus gland is a neurohemal organ, in which the neurohormones, synthesised in the neurosecretory cells of the X-organ are stored until their release. The X-organ location differs in isopods and decapods. This difference is related to the fact that the former have sessile eyes leading to a reduction of the optic medulla; as a consequence, the X-organ is shifted to the median part of the brain. However, in both groups the sinus gland neurohormones are involved in monitoring almost all known physiological processes [2]. The crustacean hyperglycemic hormone (CHH) has been shown to be the most abundant hormone in the sinus gland of isopods and decapods and controls the blood sugar level [3, 41. Molting and reproduction are regulated by inhibitory hormones : molt inhibiting hormone (MIH), vitellogenesis inhibiting hormone (VIH), and androgenic inhibiting hormone (AIH) [5]. From the results published in recent years [4, 6, 71 it could not be excluded that CHH also interacts in various physiological processes. Furthermore, CHH was also reported to act as a stress hormone [2]. From a phylogenetic point of view, a comparison of such a hormone in marine and terrestrial related forms (Crustacea) should be very fruitful. Until now very little structural work has been done on isopod sinus gland peptides. We first reported the CHH amino-acid composition [3] and, later, a partial sequence of two peptides of the CHH family [8]. Recently several papers have been published on the sinus gland peptides of decapods. These peptides exist as different isoforms but share CHH and other biological activities [9]. This paper reports for the first time the complete aminoacid sequence of an isopod CHH. It was determined by a combination of Edman degradation and mass spectrometry [lo-131. The assignment of the position of the disulfide bridges was made unambiguously by a two-step procedure which combined Edman degradation and mass spectrometry. MATERIALS AND METHODS Experimental animalsArmadillidium vulgare woodlice (C...

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“…From these triply and doubly protonatcd ions, the molccular mass of the fragment was deduced to correspond to 1558.6 Da. A computer program [16] was used to predict the molecular masses of all possible cleavage products which could be obtained with all theoretically possible combinations of disulfide bridges within the pcptidc. This program includes clcavages at all 37 peptides bonds of the antibacterial peptide and not only at the expected enzymic cleavage sites mentioned above.…”

Section: Determination Of Disulfide Bondsmentioning

confidence: 99%

A novel insect defensin mediates the inducible antibacterial activity in larvae of the dragonfly Aeschna cyanea (Paleoptera, Odonata)

Bulet

Cociancich

Reuland

et al. 1992

European Journal of Biochemistry

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114

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The injection of low doses of bacteria into the aquatic larvae of dragonflies (Aeschna cyanea, Odonata, Paleoptera) induces the appearance in their hemolymph of a potent antibacterial activity. We have isolated a 38‐residue peptide from this hemolymph which is strongly active against Gram‐positive bacteria and also shows activity against one of the Gram‐negative bacteria which was tested. The peptide is a novel member of the insect defensin family of inducible antibacterial peptides, which had so far only been reported from the higher insect orders believed to have evolved 100 million years after the Paleoptera. Aeschna defensin is more potent than defensin from the dipteran Phormia, from which its structure differs in several interesting aspects, which are discussed in the paper.

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Determination of disulfide bridges in natural and recombinant insect defensin A

Cited by 28 publications

References 16 publications

Moricin, a Novel Type of Antibacterial Peptide Isolated from the Silkworm, Bombyx mori

Moricin, a Novel Type of Antibacterial Peptide Isolated from the Silkworm, Bombyx mori

Isolation and molecular characterization of a hyperglycemic neuropeptide from the sinus gland of the terrestrial isopod Armadillidium vulgare (Crustacea)

A novel insect defensin mediates the inducible antibacterial activity in larvae of the dragonfly Aeschna cyanea (Paleoptera, Odonata)

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