{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,23]],"date-time":"2025-09-23T14:20:48Z","timestamp":1758637248793},"reference-count":27,"publisher":"FapUNIFESP (SciELO)","issue":"2","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Rev. Bras. Parasitol. Vet."],"published-print":{"date-parts":[[2014,6]]},"abstract":"<jats:p>Ticks are rich sources of serine protease inhibitors, particularly those that prevent blood clotting and inflammatory responses during blood feeding. The tick Rhipicephalus (Boophlus) annulatusis an important ectoparasite of cattle. The aims of this study were to characterize and purify the serine protease inhibitors present in R. (B.) annulatus larval extract. The inhibitors were characterized by means of one and two-dimensional reverse zymography, and purified using affinity chromatography on a trypsin-Sepharose column. The analysis on one and two-dimensional reverse zymography of the larval extract showed trypsin inhibitory activity at between 13 and 40 kDa. Through non-reducing SDS-PAGE and reverse zymography for proteins purified by trypsin-Sepharose affinity chromatography, some protein bands with molecular weights between 13 and 34 kDa were detected. Western blotting showed that five protein bands at 48, 70, 110, 130 and 250 kDa reacted positively with immune serum, whereas there was no positive reaction in the range of 13-40 kDa. Serine protease inhibitors from R. (B.) annulatus have anti-trypsin activity similar to inhibitors belonging to several other hard tick species, thus suggesting that these proteins may be useful as targets in anti-tick vaccines.<\/jats:p>","DOI":"10.1590\/s1984-29612014036","type":"journal-article","created":{"date-parts":[[2014,7,17]],"date-time":"2014-07-17T13:31:49Z","timestamp":1405603909000},"page":"187-193","source":"Crossref","is-referenced-by-count":2,"title":["Assessment and partial purification of serine protease inhibitors from Rhipicephalus (Boophilus) annulatuslarvae"],"prefix":"10.1590","volume":"23","author":[{"given":"Sedigheh","family":"Nabian","sequence":"first","affiliation":[{"name":"University of Tehran \u2013 UT; Iranian Research Center for Tick and Tick- borne Diseases"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohammad","family":"Taheri","sequence":"additional","affiliation":[{"name":"University of Tehran \u2013 UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohammad Mehdi","family":"Ranjbar","sequence":"additional","affiliation":[{"name":"University of Tehran \u2013 UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alireza","family":"Sazmand","sequence":"additional","affiliation":[{"name":"University of Veterinary Medicine Vienna \u2013 Vetmeduni Vienna; Payame Noor University \u2013 PNU"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Parastou","family":"Youssefy","sequence":"additional","affiliation":[{"name":"University of Tehran \u2013 UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gholam Reza","family":"Nazaralipour","sequence":"additional","affiliation":[{"name":"University of Tehran \u2013 UT"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"530","reference":[{"issue":"(5)","key":"ref1","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1023\/A:1012242817869","article-title":"Serine proteinase inhibitors from eggs and larvae of tick Boophilus microplus: purification and biochemical characterization","volume":"20","author":"Andreotti R","year":"2001","journal-title":"J Protein Chem"},{"issue":"(2)","key":"ref2","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1097\/00001721-199903000-00001","article-title":"Invertebrate compounds acting on the hemostatic mechanism","volume":"10","author":"Arocha-Pi\u00f1ango CL","year":"1999","journal-title":"Blood Coagul Fibrinolysis"},{"issue":"(1-2)","key":"ref3","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford MM","year":"1976","journal-title":"Ann Biochem"},{"issue":"(6)","key":"ref4","doi-asserted-by":"crossref","first-page":"3319","DOI":"10.4049\/jimmunol.133.6.3319","article-title":"Characterization of tick antigens inducing host immune resistance. I. Immunization of guinea pigs with Amblyomma americanum-derived salivary gland extracts and identification of an important salivary gland protein antigen with guinea pig anti-tick antibodies","volume":"133","author":"Brown SJ","year":"1984","journal-title":"J Immunol"},{"issue":"(9)","key":"ref5","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/j.ibmb.2009.07.003","article-title":"Tick-derived Kunitz-type inhibitors as antihemostatic factors","volume":"39","author":"Corral-Rodr\u00edguez MA","year":"2009","journal-title":"Insect Biochem Mol Biol"},{"key":"ref6","series-title":"Ticks of domestic animals in the Mediterranean region. A guide to identification of species","author":"Estrada-Pe\u00f1a A","year":"2004"},{"key":"ref7","author":"Hudson L","year":"1989"},{"issue":"(8)","key":"ref8","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1023\/A:1016136207308","article-title":"Isolation and properties of two forms of thrombin inhibitor from the nymphs of the camel tick Hyalomma dromedarii (Acari: Ixodidae)","volume":"25","author":"Ibrahim MA","year":"2001","journal-title":"Exp Appl Acarol"},{"issue":"(10)","key":"ref9","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1016\/j.vaccine.2004.08.041","article-title":"A serine protease inhibitor (serpin) fromHaemaphysalis longicornis as an anti-tick vaccine","volume":"23","author":"Imamura S","year":"2005","journal-title":"Vaccine"},{"issue":"(4-5)","key":"ref10","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S0145-305X(99)00012-9","article-title":"Serine proteinase inhibitors in arthropod immunity","volume":"23","author":"Kanost MR","year":"1999","journal-title":"Dev Comp Immunol"},{"issue":"(5259)","key":"ref11","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4","volume":"227","author":"Laemmli UK","year":"1970","journal-title":"Nature"},{"issue":"(1-2)","key":"ref12","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1016\/S0167-4838(99)00284-8","article-title":"What can the structures of enzyme-inhibitor complexes tell us about the structures of enzyme substrate complexes?","volume":"1477","author":"Laskowski M","year":"2000","journal-title":"Biochim Biophys Acta"},{"issue":"(10)","key":"ref13","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1292\/jvms.63.1063","article-title":"Tick-Encoded serine proteinase inhibitors (serpins); potential target antigens for tick vaccine development","volume":"63","author":"Muleng A","year":"2001","journal-title":"J Vet Med Sci"},{"issue":"(3)","key":"ref14","first-page":"428","article-title":"A new family of serine protease inhibitors (Bombyx family) as established from the unique topological relation between the positions of disulphide bridges and reactive site","volume":"119","author":"Pham TN","year":"1996","journal-title":"J Biochem"},{"issue":"(17)","key":"ref15","doi-asserted-by":"crossref","first-page":"3284","DOI":"10.1016\/j.vaccine.2007.01.008","article-title":"Protective immunity against Ixodes ricinus induced by a salivary serpin","volume":"25","author":"Prevot PP","year":"2007","journal-title":"Vaccine"},{"issue":"(3-4)","key":"ref16","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/j.vetpar.2008.04.020","article-title":"Serologically definedRhipicephalus (Boophilus) micropolus larval antigens in BmLF3, a partially pure Sephacryl S-300 fraction of crude larval proteins","volume":"155","author":"Pruett JH","year":"2008","journal-title":"Vet Parasitol"},{"issue":"(2)","key":"ref17","first-page":"176","article-title":"Rhipicephalus sanguineus trypsin inhibitors present in the tick larvae: isolation, characterization, and partial primary structure determination","volume":"417","author":"Sant'Anna Azzolini S","year":"2003","journal-title":"Arch Biochem Biophys"},{"issue":"(9-10)","key":"ref18","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1016\/j.biochi.2004.09.010","article-title":"Boophilus microplus tick larvae, a rich source of Kunitz type serine proteinase inhibitors","volume":"86","author":"Sasaki SD","year":"2004","journal-title":"Biochimie"},{"key":"ref19","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1146\/annurev.en.40.010195.001333","article-title":"Tick salivary gland physiology","volume":"40","author":"Sauer JR","year":"1995","journal-title":"Annu Rev Entomol"},{"issue":"(1)","key":"ref20","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S1096-4959(01)00530-9","article-title":"Structural and functional properties of a novel serine protease inhibiting peptide family in arthropods","volume":"132","author":"Simonet G","year":"2002","journal-title":"Comp Biochem Physiol B Biochem Mol Biol"},{"issue":"(1-3)","key":"ref21","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/S0162-3109(99)00074-0","article-title":"A double headed serine proteinase inhibitor--human plasma kallikrein and elastase inhibitor-- from Boophilus microplus larvae","volume":"45","author":"Tanaka AS","year":"1999","journal-title":"Immunopharmacology"},{"issue":"(6)","key":"ref22","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1016\/0020-711X(88)90102-4","article-title":"Kinetic properties of toxic protease inhibitors isolated from tick eggs","volume":"20","author":"Vermeulen NM","year":"1988","journal-title":"Int J Biochem"},{"issue":"(Pt 4)","key":"ref23","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1017\/S003118200008077X","article-title":"Comparison of the proteins in salivary glands, saliva and haemolymph of Rhipicephalus appendiculatusfemale ticks during feeding","volume":"109","author":"Wang H","year":"1994","journal-title":"Parasitology"},{"issue":"(1)","key":"ref24","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1042\/bj1770041","article-title":"Characterization of a proteolytic-enzyme inhibitor with allergenic activity. Multiple functions of a parasite-derived protein","volume":"177","author":"Willadsen P","year":"1979","journal-title":"Biochem J"},{"issue":"(Pt 2)","key":"ref25","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1038\/icb.1983.21","article-title":"Trypsin-chymotrypsin inhibitors from the tick,Boophilus microplus","volume":"61","author":"Willadsen P","year":"1983","journal-title":"Aust J Exp Biol Med Sci"},{"issue":"(19)","key":"ref26","doi-asserted-by":"crossref","first-page":"16941","DOI":"10.1074\/jbc.M212453200","article-title":"Comprehensive analysis of the secreted proteins of the parasite Haemonchus contortus reveals extensive sequence variation and differential immune recognition","volume":"278","author":"Yatsuda AP","year":"2003","journal-title":"J Biol Chem"},{"issue":"(9)","key":"ref27","doi-asserted-by":"crossref","first-page":"1970","DOI":"10.1128\/IAI.55.9.1970-1978.1987","article-title":"Isolation and characterization of an extracellular proteinase of Coccidioides immitis","volume":"55","author":"Yuan L","year":"1987","journal-title":"Infect Immun"}],"container-title":["Revista Brasileira de Parasitologia Veterin\u00e1ria"],"original-title":[],"link":[{"URL":"http:\/\/www.scielo.br\/pdf\/rbpv\/v23n2\/0103-846X-rbpv-23-02-187.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,15]],"date-time":"2023-07-15T09:08:30Z","timestamp":1689412110000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.scielo.br\/scielo.php?script=sci_arttext&pid=S1984-29612014000200187&lng=en&tlng=en"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,6]]},"references-count":27,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2014,6]]}},"alternative-id":["S1984-29612014000200187"],"URL":"https:\/\/doi.org\/10.1590\/s1984-29612014036","relation":{},"ISSN":["1984-2961"],"issn-type":[{"value":"1984-2961","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,6]]}}}