Molecular cloning of a cDNA and assignment of the <i>C</i>-terminal of sarcotoxin IA, a potent antibacterial protein of <i>Sarcophaga peregrina</i>

Matsumoto, Noriko; Okada, Mami; Takahashi, Hideo; Ming, Q X; Nakajima, Yuki; Nakanishi, Yoko; Komano, Hiroto; Natori, Shunji

doi:10.1042/bj2390717

Cited by 81 publications

(38 citation statements)

References 29 publications

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“…We assigned the signal sequence of this enzyme as 20 amino acid residues from the first Met to Ala at position 20, because this region is hydrophobic, as shown in Fig. 2, and the carboxyl terminal amino acid residues of the signal sequences of all secretory proteins of Sareophaga so far examined are Ala [13,14]. Thus the pro-segment of this enzyme may be the 67 amino acid residues from position 21 to 88.…”

Section: Sized As Ga(tic)gticciga(aig)ga(aig)tt(tic)ga(tic)gcmentioning

confidence: 99%

Molecular cloning of cDNA for the 29 kDa proteinase participating in decomposition of the larval fat body during metamorphosis of Sarcophaga peregrina (flesh fly)

1993

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A cDNA clone for the 29 kDa proteinase participating in tissue disintegration during metamorphosis of Sareophaga was isolated. This proteinase, named Sarcophaga cathepsin B, consisted of 256 amino acid residues, and contained three putative N-glycosylation sites. By comparison with other cathepsins B, its unique substrate specificity was partly explained by Ala at position 248. cDNA cloning; Cathepsin B; Metamorphosis; Fat body decomposition

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Section: Sized As Ga(tic)gticciga(aig)ga(aig)tt(tic)ga(tic)gcmentioning

confidence: 99%

Molecular cloning of cDNA for the 29 kDa proteinase participating in decomposition of the larval fat body during metamorphosis of Sarcophaga peregrina (flesh fly)

1993

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“…Furthermore, the measured mass compared to the predicted mass (4060.6 Da) from the amino acid sequence, eliminated any possibility of post-translational modifications involving variations of more than 1 Da. A monoisotopic mass measurement (measured, 4057.90 Da; expected, 4057.81 Da) with an accuracy better than 0.1 Da was also achieved [l] and it was thus possible to demonstrate that the C-terminus was not amidated (expected mass difference of -1 Da) although this has been described for other antibacterial peptides like cecropin isolated from Hyalophora cecropia [2] and the related sarcotoxin IA isolated from Sarcophaga peregrina [3], as well as diptericin isolated from P. terranovae [4]. Finally, the data obtained from mass spectrometry measurements also showed that natural insect defensin A was not present in a covalent dimeric form since the molecular ion did not exhibit peaks separated by mjz 0.5, and no peak was detected at mjz 8120 [l].…”

Section: ---mentioning

confidence: 99%

Determination of disulfide bridges in natural and recombinant insect defensin A

Lepage

Bitsch

Roecklin

et al. 1991

European Journal of Biochemistry

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The primary-structure comparison of natural insect defensin A from Phormia terranovae and recombinant insect defensin A from Saccharomyces cerevisiae has been accomplished using a combination of Edman degradation and liquid secondary ion mass spectrometry. The natural and recombinant proteins have the same primary structure with identical disulfide-bond designations (Cys3 Cys30, Cysl6 Cys36 and Cys20 Cys38) as determined from the peptides obtained after thermolysin digestion. The combined use of Edman degradation and mass spectrometry allowed the disulfide-bridge structure to be determined with a total of only 40 pg (9.9 nmol) natural peptide. Mass spectrometry provides a rapid means of disulfide-bridge verification, requiring not more than 20 pg recombinant insect defensin A, which is compatible with use in batch analysis.

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confidence: 99%

“…Working alongside Dr. Zong-Qu Li (a visitor from the University of Wuhan, in China), we prepared both precursors and their putative downstream products in suitably labeled forms that allowed to study their posttranslational processing in considerable detail. 9 By then, other laboratories, mainly Lehrer's at UCLA, 10 Shunji Natori's in Tokyo [11][12][13] 14 had discovered similar antimicrobial activities in biological systems other than Boman's Cecropia moth, and in several cases applied synthetic peptide chemistry to explore mechanisms of action and to develop analogues with improved properties. The field of antimicrobial peptide research was taking off to become one of the most dynamic areas of peptide-related research over the next decades.…”

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confidence: 99%

“…19 Downsized hybrids such as CA(1-8)M (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18) or CA(1-7)M(5-9) by way of their ready synthetic accessibility and good antimicrobial activity, have since been adopted as models in numerous studies on mechanism of action, [20][21][22][23][24][25][26] as well as explored for prospective therapeutic application to a number of targets. [27][28][29][30][31][32][33] Bruce's lab contributions to the field continued steadily well into the last decade, [34][35][36][37] mostly through the efforts of Drs.…”

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Bruce Merrifield's contribution to antimicrobial peptide research

Andreu

2008

Biopolymers

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A lthough within the scientific community at large Bruce Merrifield is and will remain celebrated for the pivotal discovery and dedicated crafting of solid phase peptide synthesis methodology, there are also other areas where his unique scientific acumen was applied to with distinctive success. Among them, those of glucagon antagonists and antimicrobial peptides are arguably the better known. Over my 25 year-long relationship with Bruce, I had occasion to work with him in both areas, but it is to the latter I will devote these brief reminiscences, as our collaboration in this fast expanding research field reached from its very dawn till relatively recently.2006, the year of Bruce's passing, marked the silver jubilee of antimicrobial peptide research, officially initiated with the publication of the seminal paper on the cecropins by Boman (see Figure 1) and coworkers. 1 Hans had befriended Bruce during his postdoc in Fritz Lipmann's group at Rockefeller, and his logical enthusiasm over the finding of a family of eukaryotic, geneencoded peptides with remarkably potent antimicrobial activities must have been contagious for Bruce, who started to collaborate with Hans on the cecropins in late 1981. On joining Bruce's lab in January 1982, the very first project given to me was to synthesize the putative 37-residue sequence of cecropin A, to confirm some structural assignations tentatively proposed for the C-terminal region. A previous synthetic attempt 2 had turned out an active compound, albeit nonidentical with the natural peptide. Building on that result and benefiting from several improvements emanating at the time from Bruce and Jimmy Tam's joint research-not least the very useful high-low HF cleavage method 3 -I completed my assignment in two-and-a-half months, which at the time was regarded a reasonable time for a relative novice charged with the fully manual synthesis of a 37-residue, tritiumlabeled peptide. Our synthetic product was completely identical with the native peptide by microbiological as well as biochemical (enzyme digest + HPLC) criteria, and we were able to confirm the hitherto uncertain structure of the C-terminal end by means of plasma desorption MS, a forerunner of modern MALDI-TOF MS. In addition, our report 4 discussed the relevance of the Trp-2 residue for antimicrobial activity and thus set the basis for further analogue work. 5 In retrospect, that initial 2-year stint in the cecropin project appears as a decisive period, where Bruce's gracious support and encouragement became the beacon blazing away the often usual angst of a nascent scientific career. A thorough and constructive draft reviewer if ever I saw one, Bruce returned each one of my versions generously endowed with pencil scribbles which, readily deciphered, brought gradual improvement to my struggling manuscripts. To Bruce I owe a great deal of any abilities I may have achieved as a (non-native) writer of English scientific texts. Those early 1980s cecropin manuscripts, first examples of the substantial contribution synthetic ...

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Molecular cloning of a cDNA and assignment of the C-terminal of sarcotoxin IA, a potent antibacterial protein of Sarcophaga peregrina

Cited by 81 publications

References 29 publications

Molecular cloning of cDNA for the 29 kDa proteinase participating in decomposition of the larval fat body during metamorphosis of Sarcophaga peregrina (flesh fly)

Molecular cloning of cDNA for the 29 kDa proteinase participating in decomposition of the larval fat body during metamorphosis of Sarcophaga peregrina (flesh fly)

Determination of disulfide bridges in natural and recombinant insect defensin A

Bruce Merrifield's contribution to antimicrobial peptide research

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