Chemical synthesis and characterization of maurocalcine, a scorpion toxin that activates Ca(2+) release channel/ryanodine receptors.
Maurocalcine is a novel toxin isolated from the venom of the chactid scorpion Scorpio maurus palmatus. It is a 33-mer basic peptide cross-linked by three disulfide bridges, which shares 82% sequence identity with imperatoxin A, a scorpion toxin from the venom of Pandinus imperator. Maurocalcine is peculiar in terms of structural properties since it does not possess any consensus motif reported so far in other scorpion toxins. Due to its low concentration in venom (0.5% of the proteins), maurocalcine was chemically synthesized by means of an optimized solid-phase method, and purified after folding/oxidation by using both C18 reversed-phase and ion exchange high-pressure liquid chromatographies. The synthetic product (sMCa) was characterized. The half-cystine pairing pattern of sMCa was identified by enzyme-based cleavage and Edman sequencing. The pairings were Cys3-Cys17, Cys10-Cys21, and Cys16-Cys32. In vivo, the sMCa was lethal to mice following intracerebroventricular inoculation (LD(50), 20 microg/mouse). In vitro, electrophysiological experiments based on recordings of single channels incorporated into planar lipid bilayers showed that sMCa potently and reversibly modifies channel gating behavior of the type 1 ryanodine receptor by inducing prominent subconductance behavior.
Fajloun
Z
Z
Laboratoire de Biochimie, CNRS UMR 6560, IFR Jean Roche, Faculté de Médecine Nord, Bd Pierre Dramard, 13916, Marseille, France. fajloun.z@jean-roche.univ-mrs.fr
Kharrat
R
R
Chen
L
L
Lecomte
C
C
Di Luccio
E
E
Bichet
D
D
El Ayeb
M
M
Rochat
H
H
Allen
P D
PD
Pessah
I N
IN
De Waard
M
M
Sabatier
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003553:Cystine / Q000737:chemistry; D004594:Electrophysiology; D007276:Injections, Intraventricular; D015640:Ion Channel Gating; D007928:Lethal Dose 50; D008051:Lipid Bilayers; D051379:Mice; D008969:Molecular Sequence Data; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry*; D012604:Scorpion Venoms / Q000138:chemical synthesis* / Q000737:chemistry / Q000633:toxicity; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid
D008051:Lipid Bilayers; D019837:Ryanodine Receptor Calcium Release Channel; D012604:Scorpion Venoms; C406406:maurocalcine; D003553:Cystine