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Snake Venom Proteins: Development Into Antimicrobial and Wound Healing Agents

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Snake Venom Proteins: Development Into Antimicrobial and Wound Healing Agents Send Orders for Reprints to [email protected] 4 Mini-Reviews in Organic Chemistry, 2014, 11, 4-14 Snake Venom Proteins: Development into Antimicrobial and Wound Healing Agents 1,2 3 4 5 Ramar Perumal Samy *, Jayapal Manikandan , Gautam Sethi , Octavio L. Franco , Josiah C. Okonkwo6, Bradley G. Stiles7,#, Vincent T.K. Chow1, Ponnampalam Gopalakrishnakone2 and Mohammed Al Qahtani3 1Infectious Diseases Programme, Department of Microbiology, 2Venom and Toxin Research Programme, Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Singapore - 117597; 3Center of Excellence in Genomic Medicine Research, King Abdulaziz University, PO Box 80216, Jeddah 21589, Kingdom of Saudi Arabia; 4Department of Pharmacology, Clinical Research Centre, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System (NUHS), Singapore – 117597; 5Universidade Católica de Brasília, Centro de Análises Proteômicas e Bioquímicas, Pós-Graduação em Ciências Genômicas e Biotecnologia UCB, Brasília-DF, Brazil; 6Department of Animal Science and Technology, Nnamdi Azikiwe University, PMB 5025 Awka, Anambra, Nigeria; 7Integrated Toxicology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 21702-5011, USA; #Department of Biology, Wilson College, 1015 Philadelphia Avenue, Chambersburg, Pennsylvania 17201, USA Abstract: Infectious diseases are a significant cause of morbidity and mortality worldwide, accounting for approximately 50% of all deaths in tropical countries and as much as 20% of deaths in the USA. The emergence of multi-drug resistant (MDR) strains makes the risk of these infections even more threatening and an important public health problem thereby increasing need of new agents for fighting pathogens. In this review, the remarkable antibacterial properties possessed by various snake venoms (Crotalide, Elapidae, and Viperidae families) were discussed and in particular phospholipase A2s (PLA2s) that have emerged from various studies as potential in the last few years. Group IIA PLA2s are the most potent among the snake venom (sv)PLA2s against various types of bacteria. Further, antibacterial derivatives from PLA2s, e.g. peptides derived from the C-terminal sequence of Lys49-PLA2s (amino acids 115-129), kill bacteria and cause severe membrane-damaging effects. Mechanisms of binding to the bacterial surface and subsequent killing by peptides are based on positive charge, hydrophobicity, and length. These peptide candidates are easy to design and synthesize in pure form (~95% purity). Such peptides may be potentially useful in the clinic as new antimicrobials for combating infections due to antibiotic-resistant bacteria that include methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus in the near future. Keywords: Methicillin-resistant Staphylococcus aureus (MRSA), snake venom protein, endogenous antibiotics, Lys49 & Asp49-PLA2, inflammatory cytokines, growth factors, skin, wound healing, transcription factors NF-kB, Cys-rich protein. 1. INTRODUCTION infections. SA pneumonia has increased in frequency over the years and most have been caused by community-acquired Scabies cause discomfort to the patient and serious MRSA and in particular USA 300 isolates. Viral co-infection complication as a results of secondary bacterial infection with S. aureus in 15% of the cases is associated with caused by Streptococcus pyogenes (GAS) or Staphylococcus respiratory failure [4]. Bacterial infections involving multi- aureus [1]. Recently, a high burden of skin and soft tissue drug-resistant (MDR) strains are one of the ten leading infections (a total of 7252 isolates), and 55% of methicillin- causes of death world-wide according to the WHO [5]. The resistant S. aureus (MRSA) bloodstream infections were emergence of MDR strains makes the risk of these infections reported and spread out in hospitals across different regions more threatening now [6] and an important health problem in of the world. [2]. Particularly, cellulitis (up to 38% of need of new antibacterial strategies. Besides, antimicrobial patients affected by these diseases) is an important serious resistance is recognized as one of the greatest threats to skin infections and increasing problem in children [3]. The human health in worldwide [7]. The reasons for this rate of community-acquired S. aureus (SA) pneumonia per resistance include a reduction in the new antibacterial drugs 10,000 admissions increased from 4.81% hospitalizations in in the pharmaceutical pipeline, an increase in antimicrobial year 2001 to 2009. MRSA caused 74% and methicillin- resistance, and the need of treatments for newly discovered susceptible (MSSA) caused 26% of these pneumonia pathogens. Venoms from snakes in the Viperidae, Crotadiae and *Address correspondence to this author at the Venom and Toxin Research Elapidae families are known to consist of complex mixtures Programme, Department of Anatomy, Yong Loo Lin School of Medicine, of proteins and peptides responsible for various toxic effects. National University of Singapore, Singapore 117597; Tel: 65-65168733; Fax: +65-67787643; E-mail: [email protected] Snake venom contains numerous components of biological 1875-6298/14 $58.00+.00 © 2014 Bentham Science Publishers Snake Venom Proteins: Development into Antimicrobial and Wound Healing Agents Mini-Reviews in Organic Chemistry, 2014, Vol. 11, No. 1 5 Vancomycin Quinolone Tetracycline Cell wall synthesis Protein DNA (A) synthesis gyrase Bacteria Sulfanomide Folic acid DNA directed RNA metabolism polymorphism Susceptible Bacteria Rifampicin Mutation Foreign DNA (Chromosomal) (Acquisition) Antibiotic Selection Pressure Aminoglycosides by enzyme by Modification Inactivation/ Antibiotic Target mutation Efflux pump Reduced permeability Resistant Bacteria Imipenem Tetracycline Rifampicin Fig. (1). Possible mechanisms of susceptibility by various antibiotics and representation of the emergence of drug-resistant bacteria. interest. Venom is a complex mixture that varies in addition, PLA2s can have a significant antibacterial effect not proportions and characteristics among different species, strictly dependent on its enzymatic activity [19]. Furthe- containing non-protein and protein components with varying rmore, a PLA2-derived peptide exhibits fungicidal activity structures and biochemical activities. Major protein com- against Candida albicans [20]. An antimicrobial peptide ponents are neurotoxins, cardiotoxins, lectins, disintegrins, derived from the C-terminus of (B. asper) myotoxin II natriuretic peptides, proteases, phospholipases, phospho- interacts with phospholipid bilayers [21]. Another report diesterases, nucleotidases, and L-amino acid oxidases [8]. In shows that cationic peptides designed from a Lys49 PLA2 of addition, venom enzymes exert a variety of pharmacological snake venom possesses bactericidal and anti-endotoxic effects such as neurotoxicity, myotoxicity, cardiotoxicity, properties [22]. However, those peptides (amino acids 115- hemorrhage, pro- and anti-coagulation, etc. [9, 10]. Venoms 129) derived from the carboxy terminus are devoid of and more specifically their phospholipase A2s (PLA2s) have toxicity and are more bactericidal than the parent molecules. demonstrated antibacterial activity [11-13]. Type-IIA These peptide derivatives of snake venom (sv)PLA2s, and secretory (s) PLA2 is an endogenous antibiotic-like protein other biologically active proteins from venoms, may lead to of the host [14-17]. Lys49 and Asp49 myotoxic PLA2s from the production of new drugs of potential therapeutic value in Bothrops asper also possess bactericidal activity [18]. In the near future (Fig. 1) [23, 24]. 6 Mini-Reviews in Organic Chemistry, 2014, Vol. 11, No. 1 Samy et al. 2. BIOLOGICAL AND CELLULAR FUNCTIONS OF events regulating cellular functions such as direct killing svPLA2 of bacteria and modulation of inflammatory responses (Fig. 2A) [33]. Most PLA2s catalyze hydrolysis of glycerophospholipids at the sn-2 position to release free fatty acids and lysophos- 4. BACTERICIDAL ACTIVITY OF SNAKE VENOMS pholipids, that plays an important role in biosynthesis of biologically active lipid mediators [25]. A recent study Antimicrobial agents are used to treat infections by reported a variety of membrane and soluble proteins that microbes, which include different classes of pathogenic bind to sPLA2s, suggesting that sPLA2s may also function as organisms-viruses, rickettsia, mycoplasma, chlamydia, high affinity ligands [26]. Collectively, venom-derived protozoa, bacteria and fungi. Bacteria are the largest and sPLA2s bind to membrane and soluble mammalian proteins most diverse group of pathogenic microorganisms [34]. that include the C-type lectin superfamily (M-type sPLA2 There are a number of antimicrobial agents normally used to receptors and surfactant proteins), pentraxin and reticulo- treat infections caused by bacteria, on which they have a calbin, factor Xa, and N-type receptors [25]. Otherwise, selective toxic action. Bacteriostatic agents act primarily by mammalian group IB and IIA sPLA2s can bind to the M-type arresting growth (e.g. sulphonamides, tetracycline, chlora- receptor, and group IIA sPLA2 can associate with lung mphenicol), as compared to the bactericidal agents, which surface proteins, factor Xa and proteoglycans including act primarily by killing cells through disruption of cell glypican and decorin, a mammalian protein containing a wall/membrane
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