Crystallization and preliminary X-ray diffraction studies of La1 from<i>Liocheles australasiae</i>

Kamachi, Saori; Nagao, Junya; Miyashita, Masahiro; Nakagawa, Yoshiaki; Miyagawa, Hisashi; Tada, Toshiji

doi:10.1107/s2053230x14010589

Cited by 2 publications

(2 citation statements)

References 16 publications

(19 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The La1-like peptides are presumed to possess a single Von Willebrand factor type C domain (VWC), a structural feature shared by several eukaryotic proteins with cell biology functions [ 75 , 76 ], although this has not been proven [ 77 ].…”

Section: Resultsmentioning

confidence: 99%

Transcriptome Analysis of Scorpion Species Belonging to the Vaejovis Genus

et al. 2015

View full text Add to dashboard Cite

Scorpions belonging to the Buthidae family have traditionally drawn much of the biochemist’s attention due to the strong toxicity of their venoms. Scorpions not toxic to mammals, however, also have complex venoms. They have been shown to be an important source of bioactive peptides, some of them identified as potential drug candidates for the treatment of several emerging diseases and conditions. It is therefore important to characterize the large diversity of components found in the non-Buthidae venoms. As a contribution to this goal, this manuscript reports the construction and characterization of cDNA libraries from four scorpion species belonging to the Vaejovis genus of the Vaejovidae family: Vaejovis mexicanus, V. intrepidus, V. subcristatus and V. punctatus. Some sequences coding for channel-acting toxins were found, as expected, but the main transcribed genes in the glands actively producing venom were those coding for non disulfide-bridged peptides. The ESTs coding for putative channel-acting toxins, corresponded to sodium channel β toxins, to members of the potassium channel-acting α or κ families, and to calcium channel-acting toxins of the calcin family. Transcripts for scorpine-like peptides of two different lengths were found, with some of the species coding for the two kinds. One sequence coding for La1-like peptides, of yet unknown function, was found for each species. Finally, the most abundant transcripts corresponded to peptides belonging to the long chain multifunctional NDBP-2 family and to the short antimicrobials of the NDBP-4 family. This apparent venom composition is in correspondence with the data obtained to date for other non-Buthidae species. Our study constitutes the first approach to the characterization of the venom gland transcriptome for scorpion species belonging to the Vaejovidae family.

show abstract

Section: Resultsmentioning

confidence: 99%

Transcriptome Analysis of Scorpion Species Belonging to the Vaejovis Genus

et al. 2015

View full text Add to dashboard Cite

show abstract

“…This suggests that La1 has similar conformation to the VWC domain in ECM proteins. Currently, X-ray structural analysis of La1 is in progress to confirm this hypothesis [19].…”

Section: Determination Of Disulfide Bonding Patternmentioning

confidence: 97%

Chemical synthesis of La1 isolated from the venom of the scorpion Liocheles australasiae and determination of its disulfide bonding pattern

Nagao

Miyashita

Nakagawa

et al. 2015

Journal of Peptide Science

Self Cite

View full text Add to dashboard Cite

La1 is a 73-residue cysteine-rich peptide isolated from the scorpion Liocheles australasiae venom. Although La1 is the most abundant peptide in the venom, its biological function remains unknown. Here, we describe a method for efficient chemical synthesis of La1 using the native chemical ligation (NCL) strategy, in which three peptide components of less than 40 residues were sequentially ligated. The peptide thioester necessary for NCL was synthesized using an aromatic N-acylurea approach with Fmoc-SPPS. After completion of sequential NCL, disulfide bond formation was carried out using a dialysis method, in which the linear peptide dissolved in an acidic solution was dialyzed against a slightly alkaline buffer to obtain correctly folded La1. Next, we determined the disulfide bonding pattern of La1. Enzymatic and chemical digests of La1 without reduction of disulfide bonds were analyzed by liquid chromatography/mass spectrometry (LC/MS), which revealed two of four disulfide bond linkages. The remaining two linkages were assigned based on MS/MS analysis of a peptide fragment containing two disulfide bonds. Consequently, the disulfide bonding pattern of La1 was found to be similar to that of a von Willebrand factor type C (VWC) domain. To our knowledge, this is the first report of the experimental determination of the disulfide bonding pattern of peptides having a single VWC domain as well as their chemical synthesis. La1 synthesized in this study will be useful for investigation of its biological role in the venom.

show abstract

Crystallization and preliminary X-ray diffraction studies of La1 fromLiocheles australasiae

Cited by 2 publications

References 16 publications

Transcriptome Analysis of Scorpion Species Belonging to the Vaejovis Genus

Transcriptome Analysis of Scorpion Species Belonging to the Vaejovis Genus

Chemical synthesis of La1 isolated from the venom of the scorpion Liocheles australasiae and determination of its disulfide bonding pattern

Contact Info

Product

Resources

About