Acta Biologica Hungarica 69(1), pp. 110–113 (2018) DOI: 10.1556/018.68.2018.1.9
SMALLEST LECTIN-LIKE PEPTIDE IDENTIFIED FROM THE SKIN SECRETION OF AN ENDEMIC FROG, HYDROPHYLAX BAHUVISTARA
SHORT COMMUNICATION
THUNDIPARAMPIL VASANTH VINEETHKUMAR, GOPAL SHYLA and6$1,/*(25*(
Chemical and Environmental Biology group, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram-695014, Kerala, India
(Received: September 14, 2017; accepted: December 27, 2017)
Lectins are sugar-binding proteins and considered as attractive candidates for drug delivery and targeting. +HUHZHUHSRUWWKHLGHQWL¿FDWLRQRIWKHVPDOOHVWOHFWLQOLNHSHSWLGH RGRUUDQDOHFWLQ+ Keywords: Agglutination – amphibian – antimicrobial – Hydrophylax – lectin /HFWLQV DUH SURWHLQV WKDW FDQ ELQG WR VSHFL¿F VXJDU UHVLGXHV DQG DJJOXWLQDWH FHOOV Typically they bind to cell surface glycoproteins and glycolipids [4]. Their binding is UREXVW UDSLG DQG LV GHWHUPLQHG E\ VSHFL¿F VXJDU FRGH /HFWLQV ZHUH ¿UVW LVRODWHG from plants and thought to be only an agglutinating agent. As lectins were proved to be useful tools for the investigation of sugar moieties on the cell surface, especially on cancerous cells and as mediators for drug targeting, many lectins were reported from microorganisms and animals [4]. Currently, lectins are being increasingly inves- tigated for drug targeting, which could prevent side effects to normal cells and enhance drug delivery to targeted cells [3]. The size of the lectin (more than 10 KDa) is one of the major problems which results in toxicity and immunogenicity and hence, VKRUWHUOHFWLQOLNHPROHFXOHVDUHDWWKHIRUHIURQWRIWKHKXQW>@&DWLRQLFKRVWGHIHQVH SHSWLGHV +'3 IRXQG LQ WKH VNLQ VHFUHWLRQ RI IURJV DFW DV WKH ¿UVW OLQH LPPXQH defense protecting the animal from microbial infections and predators [7]. Lectin is one such a peptide that can agglutinate bacteria and fungus and is considered as a novel defense mechanism by amphibian immune system [2]. ,QWKHSUHVHQWVWXG\DQRYHOOHFWLQOLNHSHSWLGHKDYLQJKRPRORJ\WRDQRGRUUDQD- OHFWLQIDPLO\RISHSWLGHVZDVLGHQWL¿HGIURPWKHVNLQVHFUHWLRQRIHydrophylax bahu- vistaraDQHQGHPLFIURJVSHFLHVRI:HVWHUQ*KDWV,QGLD6NLQVHFUHWLRQKDUYHVWLQJ &RUUHVSRQGLQJDXWKRUHPDLODGGUHVVVJHRUJH#UJFEUHVLQ 0236-5383/$ 20.00 © 2018 Akadémiai Kiadó, Budapest Shortest lectin-like peptide from an Indian frog 111 molecular cloning and primary structure elucidation of the peptides were done as per VWDQGDUG SURWRFROV >@ +RPRORJ\ VHDUFK RI WKH SHSWLGH VHTXHQFH ZDV GRQH XVLQJ BLAST (NCBI), Peptide was chemically synthesized by Fmoc chemistry using CLEAR-amide resin, purity of the synthesized peptide were analyzed by MALDI- TOF-MS. ProtParm (http://expasy.org/tools/protparam.html) and Pepcalc (http:// pepcalc.com/) were used to compute physicochemical properties. Antimicrobial and hemagglutinating activity were determined as per standard procedures [2, 5]. F'1$ VHTXHQFH HQFRGLQJ WKH SHSWLGH ZLWK LGHQWLW\ WR RGRUUDQDOHFWLQ UHVLGXHV*HQ%DQNDFFQR$%: UHSRUWHGIURPOdorrana grahami [5] was LGHQWL¿HG,WVRSHQUHDGLQJIUDPHHQFRGHVDSRO\SHSWLGHFRPSRVHGRIDPLQRDFLGV RUJDQL]HG TXLWH VLPLODUO\ WR DPSKLELDQ DQWLPLFURELDO SHSWLGHV ,W FRQVLVWV RI WKUHH UHJLRQVDQ1WHUPLQDOVLJQDOSHSWLGHVHTXHQFH UHVLGXHV IROORZHGE\DQDFLGLF VSDFHU UHVLGXHV DQG WKH &WHUPLQDO PDWXUH VHTXHQFH UHVLGXHV )LJ $ There is a dibasic cleavage site (K43-R44) between the acidic spacer and mature SHSWLGHZKLFKLVEHOLHYHGWREHFOHDYHGE\WU\SVLQOLNHSURWHDVHV7KHSHSWLGHVKDUHV DVLPLODUVLJQDOSHSWLGHKRPRORJ\ZLWKWKHDQWLPLFURELDOSHSWLGHOLYLGLQ+ Fig. 1. $ 7KHSUHFXUVRURIRGRUUDQDOHFWLQ+ Acta Biologica Hungarica 69, 2018 112 THUNDIPARAMPIL VASANTH VINETHKUMAR et al. mass 1640.94 Observed 1640.89 H. bahuvistara μ M 1 QRI 8.75 NA* NA* NA* NA* NA* Theoretical PI Theoretical mass 2GRUUDQDOHFWLQ+ Table 1 Table –0.353 *5$9< Gram-positive bacteria AGAINST Gram-negative bacteria BA PEPTIDE μ M) OF HY +2 Net charge (MIC) OF PEPTIDE THE MIC ( μ M) ODORRANALECTIN 'HWDLOVRIOHFWLQOLNHSHSWLGHLGHQWL¿HGIURPWKHVNLQVHFUHWLR OF OF 15 acids CONCENTRATIONS μ M) MTCC 2695 PROFILE Number of amino MTCC 9542 PROPERTIES MTCC 497 NHIBITORY I + Acta Biologica Hungarica 69, 2018 Shortest lectin-like peptide from an Indian frog 113 200 μM. (Table 1B). The same observation was previously reported for odorranalec- tin from O. grahami >@ 2GRUUDQDOHFWLQ+ ACKNOWLEDGEMENT .6&67(.HUDODLVDFNQRZOHGJHGIRU¿QDQFLDODVVLVWDQFHDQG.HUDOD)RUHVW'HSDUWPHQWIRUVDPSOLQJ permissions. REFERENCES 1. Bies, C., Lehr, C. M., Woodley, J. F. (2004) Lectin-mediated drug targeting: history and applications. Drug Deliv. Rev. 56, 425–435. /L-:X++RQJ-;X; Acta Biologica Hungarica 69, 2018