The first serine protease inhibitor from Lasiodora sp. (Araneae: Theraphosidae) hemocytes

Abstract

This work reports, for the first time, the purification, characterization and antibacterial activity of an elastase inhibitor from Lasiodora sp. hemocytes (EILaH). The hemocyte extract inhibited chymotrypsin (22%), trypsin (44%), tissue plasminogen activator (52%), urokinase (58%) and human neutrophil elastase (99%). EILaH was purified by Trypsin-Sepharose column and RP-HPLC. SDS-PAGE of EILaH revealed a molecular mass of 8 kDa and MALDI-TOF mass spectrometry revealed a single molecular mass of 8274 Da. The amino terminal sequence determined was LPC(PF)PYQQELTC. The dissociation constant (K i) for human neutrophil elastase was 0.32 nM. Hemocyte extract exerted antibacterial effect on Bacillus subtilis and Enterococcus faecalis, while EILaH was only active against E. faecalis. Currently, Lasiodora sp. is undergoing a systematic review and this study contributes to molecular characterization of the genus. In addition, the results suggest that serine protease inhibitors expressed in Lasiodora sp. hemocytes may be involved in the defense against bacterial infection.

Biochemical and Functional Characterization ofParawixia bistriataSpider Venom with Potential Proteolytic and Larvicidal Activities

“…The P4 fraction contains the protease with greatest proteolytic activity and the highest degree of purification measured in this study, and we suggest that it belongs to the same class of metalloproteases with molecular weight in the range of 40–60 kDa. Proteases are present in the digestive fluids of different species of spiders, just as protease inhibitors have been reported and characterized in several studies [ 56 – 59 ].…”

Brazilian Theraphosidae: a toxicological point of view

“…hemocyte extract against B. subtilis (ATCC-6633), S. aureus (ATCC-6538), E. faecalis (ATCC-6057), E. coli (ATCC-25922) and K. pneumoniae (ATCC-29665) as seen in Table 2. EILaH only showed activity against E. faecalis (MIC = 227.5 µg/mL), while the hemocyte extract presented activity against B. subtilis and E. faecalis (MICs were not present by authors), indicating the presence of other antimicrobial agents in the hemocytes [49].…”

“…Soares et al [49] reported in 2011, the purification and characterization of the first serine protease inhibitor extracted from Lasiodora sp. hemocytes, which was named EILaH.…”

Antimicrobial potential of Indian cooperative spider, Stegodyphus sarasinorum, and Golden orb spider, Nephila pilipes hemolymph against pathogenic bacteria

“…The total protein content of hemolymph was represented in mg/mL. The quantification was done using a spectrophotometer at 595 nm (UNICO Spectrophotometer, SP2100 UV, China) [38].…”