{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T11:19:29Z","timestamp":1763551169440,"version":"build-2065373602"},"reference-count":28,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,3,1]],"date-time":"2024-03-01T00:00:00Z","timestamp":1709251200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI","award":["JP12672137","JP10672074"],"award-info":[{"award-number":["JP12672137","JP10672074"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Snakes contain three types of phospholipase A2 (PLA2)-inhibitory proteins in their blood, PLI\u03b1, \u03b2, and \u03b3, which protect them from their own venom, PLA2. PLI\u03b2 is the snake ortholog of leucine-rich \u03b12 glycoprotein (LRG). Since autologous cytochrome c (Cyt c) serves as an endogenous ligand for LRG, in this study, we purified snake LRGs from various snake serum samples using Cyt c affinity chromatography. All purified snake LRGs were found to be dimers linked by disulfide bonds. Laticauda semifasciata and Naja kaouthia LRGs showed no inhibitory activity against L. semifasciata PLA2 and weak inhibitory activity against Gloydius brevicauda basic PLA2. Elaphe climacophora PLI\u03b2 had weaker inhibitory activity against G. brevicauda basic PLA2 than G. brevicauda and Elaphe quadrivirgata PLIs, which are abundant in blood and known to neutralize G. brevicauda basic PLA2. Protobothrops flavoviridis LRG showed no inhibitory activity against basic venom PLA2, PL-X, or G. brevicauda basic PLA2. Binding analysis of P. flavoviridis LRG using surface plasmon resonance showed very strong binding to snake Cyt c, followed by that to horse Cyt c, weak binding to yeast Cyt c, and no binding to P. flavoviridis PL-X or BPI\/II. We also deduced the amino acid sequences of L. semifasciata and P. flavoviridis LRG by means of cDNA sequencing and compared them with those of other known sequences of PLIs and LRGs. This study concluded that snake LRG can potentially inhibit basic PLA2, but, whether it actually functions as a PLA2-inhibitory protein, PLI\u03b2, depends on the snake.<\/jats:p>","DOI":"10.3390\/toxins16030126","type":"journal-article","created":{"date-parts":[[2024,3,1]],"date-time":"2024-03-01T03:31:23Z","timestamp":1709263883000},"page":"126","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["One-Step Affinity Purification of Leucine-Rich \u03b12-Glycoproteins from Snake Sera and Characterization of Their Phospholipase A2-Inhibitory Activities as \u03b2-Type Phospholipase A2 Inhibitors"],"prefix":"10.3390","volume":"16","author":[{"given":"Ryoichi","family":"Shirai","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, Takatsuki 569-1094, Osaka, Japan"},{"name":"Misasa Onsen Hospital, Misasa 682-1097, Tottori, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kana","family":"Shibata","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, Takatsuki 569-1094, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0349-8919","authenticated-orcid":false,"given":"Shinobu","family":"Fujii","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, Takatsuki 569-1094, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8972-8863","authenticated-orcid":false,"given":"Rikiro","family":"Fukunaga","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, Takatsuki 569-1094, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seiji","family":"Inoue","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, Takatsuki 569-1094, Osaka, Japan"},{"name":"Center for the Advancement of Pharmaceutical Education, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, Takatsuki 569-1094, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1042\/bj3250527","article-title":"Purification and characterization of three distinct types of phospholipase A2 inhibitors from the blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus","volume":"325","author":"Ohkura","year":"1997","journal-title":"Biochem. 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