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Host blood protein digestion, essential for tick development and reproduction, occurs in tick midgut digestive cells driven by cathepsin proteases. Little is known about the regulation of the digestive proteolytic machinery of I. ricinus. Here we characterize a novel cystatin-type protease inhibitor, mialostatin, from the I. ricinus midgut. Blood feeding rapidly induced mialostatin expression in the gut, which continued after tick detachment. Recombinant mialostatin inhibited a number of I. ricinus digestive cysteine cathepsins, with the greatest potency observed against cathepsin L isoforms, with which it co-localized in midgut digestive cells. The crystal structure of mialostatin was determined at 1.55 \u00c5 to explain its unique inhibitory specificity. Finally, mialostatin effectively blocked in vitro proteolysis of blood proteins by midgut cysteine cathepsins. Mialostatin is likely to be involved in the regulation of gut-associated proteolytic pathways, making midgut cystatins promising targets for tick control strategies.<\/jats:p>","DOI":"10.3390\/ijms22105371","type":"journal-article","created":{"date-parts":[[2021,5,20]],"date-time":"2021-05-20T06:13:45Z","timestamp":1621491225000},"page":"5371","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Mialostatin, a Novel Midgut Cystatin from Ixodes ricinus Ticks: Crystal Structure and Regulation of Host Blood Digestion"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5644-1468","authenticated-orcid":false,"given":"Jan","family":"Kot\u00e1l","sequence":"first","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Brani\u0161ovsk\u00e1 1160\/31, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"},{"name":"Department of Medical Biology, Faculty of Science, University of South Bohemia in \u010cesk\u00e9 Bud\u011bjovice, Brani\u0161ovsk\u00e1 1760c, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4750-5992","authenticated-orcid":false,"given":"Michal","family":"Bu\u0161a","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo n. 2, 16610 Praha, Czech Republic"},{"name":"Department of Biochemistry, Faculty of Science, Charles University, Hlavova 2030\/8, 12800 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veronika","family":"Urbanov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Brani\u0161ovsk\u00e1 1160\/31, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavl\u00edna","family":"\u0158ez\u00e1\u010dov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo n. 2, 16610 Praha, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jind\u0159ich","family":"Chmela\u0159","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Science, University of South Bohemia in \u010cesk\u00e9 Bud\u011bjovice, Brani\u0161ovsk\u00e1 1760c, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5123-9017","authenticated-orcid":false,"given":"Helena","family":"Langhansov\u00e1","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Science, University of South Bohemia in \u010cesk\u00e9 Bud\u011bjovice, Brani\u0161ovsk\u00e1 1760c, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Sojka","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Brani\u0161ovsk\u00e1 1160\/31, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Mare\u0161","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo n. 2, 16610 Praha, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7526-1876","authenticated-orcid":false,"given":"Michail","family":"Kotsyfakis","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Brani\u0161ovsk\u00e1 1160\/31, 37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1289\/ehp.00108119","article-title":"Impact of climatic change on the northern latitude limit and population density of the disease-transmitting European tick Ixodes ricinus","volume":"108","author":"Lindgren","year":"2000","journal-title":"Environ. 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