Characteristics and Lethality of a Novel Recombinant  Dermonecrotic Venom Phospholipase D from  Hemiscorpius lepturus.
Hemoscorpius lepturus is the most medically important scorpion in Iran. The clinical signs of H. lepturus envenomation are remarkably similar to those reported for brown spiders, including dermonecrosis, hematuria, renal failure and even death. The lethality and toxicity of brown spiders' venom have been attributed to its phospholipase D activity. This study aims to identify a phospholipase D with possible lethality and dermonecrotic activity in H. lepturus venom. In this study, a cDNA library of the venom glands was generated by Illumina RNA sequencing. Phospholipase D (PLD) from H. lepturus was characterized according to its significant similarity with PLDs from brown spiders. The main chain designated as Hl-RecPLD1 (the first recombinant isoform of H. lepturus PLD) was cloned, expressed and purified. Sphingomyelinase, dermonecrotic and lethal activities were examined. Hl-PLD1 showed remarkable sequence similarity and structural homology with PLDs of brown spiders. The conformation of Hl-PLD1 was predicted as a "TIM beta/alpha-barrel". The lethal dose 50 (LD50) and dermonecrotic activities of Hl-RecPLD1 were determined as 3.1 μg/mouse and 0.7 cm2 at 1 μg respectively. It is the first report indicating that a similar molecular evolutionary mechanism has occurred in both American brown spiders and this Iranian scorpion. In conclusion, Hl-RecPLD1 is a highly active phospholipase D, which would be considered as the lethal dermonecrotic toxin in H. lepturus venom.
Venom and Biotherapeutics Molecules Lab., Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. e.torabi88@gmail.com.
Elham
E
Torabi
Venom and Biotherapeutics Molecules Lab., Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. behdani73042@yahoo.com.
Mahdi
M
Behdani
Venom and Biotherapeutics Molecules Lab., Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. hoseininejad62@gmail.com.
Mohammad Hosseininejad
MH
Chafi
Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. r.moazzami@gmail.com.
Reza
R
Moazzami
INSERM UMRs 1097, Aix-Marseille Université, Parc Scientifique et Technologique de Luminy, Marseille 13288, France. sabatier.jm1@libertysurf.fr.
Jean-Marc
JM
Sabatier
Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. v_khalaj@yahoo.com.
Vahid
V
Khalaj
Venom and Biotherapeutics Molecules Lab., Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. shahbazzadeh@yahoo.com.
Delavar
D
Shahbazzadeh
Venom and Biotherapeutics Molecules Lab., Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran. k_bagheri@pasteur.ac.ir.
Kamran Pooshang
KP
Bagheri
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D001483:Base Sequence; D005260:Female; D006461:Hemolysis / Q000187:drug effects; D006736:Horses; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007928:Lethal Dose 50; D008297:Male; D008807:Mice, Inbred BALB C; D010739:Phospholipase D / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D012604:Scorpion Venoms / Q000276:immunology; D012605:Scorpions; D012867:Skin / Q000187:drug effects; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism
D000906:Antibodies; D060829:Arthropod Proteins; D007106:Immune Sera; D011994:Recombinant Proteins; D012604:Scorpion Venoms; D013108:Sphingomyelin Phosphodiesterase; D010739:Phospholipase D
Hemiscorpius  lepturus;   Loxoscelidae family;   brown  spiders;   homology;   phospholipase  D;   scorpion;  dermonecrotic activity;  lethality