title: Purification and characterisation of a pronase-inducible lectin isolated from human serum.
abstract: A new lectin was purified to electrophoretic homogeneity from pronase treated human serum by a single-step of affinity chromatography on concanavalin A-Sepharose 4B. The isolated lectin agglutinated five types of vertebrate RBC, with highest titer against hen RBC. This activity was independent of divalent cations, insensitive to EDTA and specific to mannosamine, glucosamine as well as galactosamine. Purified lectin gave a single symmetrical peak in its native form with a molecular mass estimate of 6kDa in FPLC analysis and 6.5kDa by MALDI-TOF MS. SDS-PAGE analysis of the lectin revealed that it is a homo-oligomer of a 3kDa subunit protein. Isolated lectin did possess both, hemagglutinating and phenoloxidase activities, but did not exhibit any antibacterial or antifungal activities. In addition, this lectin could oxidize all nine different phenolic substrates tested, with hydroquinone proving to be the best among them. Phenoloxidase inhibitors namely, phenylthiourea and tropolone inhibited this oxidation activity.
authors.affiliation: Department of Biochemistry, Annai Veilankanni's College for Women, Chennai, 600015, India. Electronic address: beulaja@gmail.com.
authors.forename: Beulaja
authors.initials: B
authors.lastname: Manikandan
authors.affiliation: Department of Zoology, University of Madras, Guindy Campus, Chennai, 600 025, India.
authors.forename: Manikandan
authors.initials: M
authors.lastname: Ramar
authors.affiliation: Department of Biotechnology, School of Chemical and Biotechnology, SASTRA University, Thanjavur, 613 401, India.
authors.forename: Thiagarajan
authors.initials: T
authors.lastname: Raman
authors.affiliation: Department of Zoology, University of Madras, Guindy Campus, Chennai, 600 025, India.
authors.forename: Mullainadhan
authors.initials: M
authors.lastname: Periasamy
authors.affiliation: Department of Zoology, University of Madras, Guindy Campus, Chennai, 600 025, India.
authors.forename: Arumugam
authors.initials: A
authors.lastname: Munusamy
mesh_terms: D000327:Adsorption; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D002846:Chromatography, Affinity; D004492:Edetic Acid / Q000737:chemistry; D004586:Electrophoresis; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D037102:Lectins / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D014442:Monophenol Monooxygenase / Q000378:metabolism; D010670:Phenylthiourea / Q000494:pharmacology; D011402:Pronase / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity; D014334:Tropolone / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D037102:Lectins; D010670:Phenylthiourea; D014334:Tropolone; D004492:Edetic Acid; D014442:Monophenol Monooxygenase; D011402:Pronase
keywords: Characterisation; Human serum; Isolation; Lectin; Phenoloxidase; Pronase