title: Characterisation of a New Fungal Immunomodulatory Protein from Tiger Milk mushroom, Lignosus rhinocerotis.
abstract: Lignosus rhinocerotis (Tiger milk mushroom) is an important folk medicine for indigenous peoples in Southeast Asia. We previously reported its de novo assembled 34.3 Mb genome encoding a repertoire of proteins including a putative bioactive fungal immunomodulatory protein. Here we report the cDNA of this new member (FIP-Lrh) with a homology range of 54-64% to FIPs from other mushroom species, the closest is with FIP-glu (LZ-8) (64%) from Ganoderma lucidum. The FIP-Lrh of 112 amino acids (12.59 kDa) has a relatively hydrophobic N-terminal. Its predicted 3-dimensional model has identical folding patterns to FIP-fve and contains a partially conserved and more positively charged carbohydrates binding pocket. Docking predictions of FIP-Lrh on 14 glycans commonly found on cellular surfaces showed the best binding energy of -3.98 kcal/mol to N-acetylgalactosamine and N-acetylglucosamine. Overexpression of a 14.9 kDa soluble 6xHisFIP-Lrh was achieved in pET-28a(+)/BL21 and the purified recombinant protein was sequence verified by LC-MS/MS (QTOF) analysis. The ability to haemagglutinate both mouse and human blood at concentration ≥0.34 μM, further demonstrated its lectin nature. In addition, the cytotoxic effect of 6xHisFIP-Lrh on MCF-7, HeLa and A549 cancer cell lines was detected at IC50 of 0.34 μM, 0.58 μM and 0.60 μM, respectively.
authors.affiliation: Faculty of Applied Sciences, No. 1, Jalan Menara Gading, UCSI Heights, Cheras 56000 Kuala Lumpur, Malaysia.
authors.forename: V
authors.initials: V
authors.lastname: Pushparajah
authors.affiliation: Faculty of Pharmacy, Quest International University Perak (QIUP), No. 227, Plaza Teh Teng Seng (Level 2), Jalan Raja Permaisuri Bainun, 30250 Ipoh, Perak Darul Ridzuan, Malaysia.
authors.forename: A
authors.initials: A
authors.lastname: Fatima
authors.affiliation: Faculty of Applied Sciences, No. 1, Jalan Menara Gading, UCSI Heights, Cheras 56000 Kuala Lumpur, Malaysia.
authors.forename: C H
authors.initials: CH
authors.lastname: Chong
authors.affiliation: Faculty of Applied Sciences, No. 1, Jalan Menara Gading, UCSI Heights, Cheras 56000 Kuala Lumpur, Malaysia.
authors.forename: T Z
authors.initials: TZ
authors.lastname: Gambule
authors.affiliation: Faculty of Applied Sciences, No. 1, Jalan Menara Gading, UCSI Heights, Cheras 56000 Kuala Lumpur, Malaysia.
authors.forename: C J
authors.initials: CJ
authors.lastname: Chan
authors.affiliation: Ligno Biotech Sdn. Bhd. No. 1, Jalan Perindustrian Balakong Jaya 2/2, Taman Perindustrian Balakong Jaya 2, 43300 Balakong Jaya, Selangor, Malaysia.
authors.forename: S T
authors.initials: ST
authors.lastname: Ng
authors.affiliation: Horticulture Research Center, MARDI Headquarters, Serdang, P.O. Box 12301, 50774 Kuala Lumpur, Malaysia.
authors.forename: C S
authors.initials: CS
authors.lastname: Tan
authors.affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: S Y
authors.initials: SY
authors.lastname: Fung
authors.affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: S S
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: N H
authors.initials: NH
authors.lastname: Tan
authors.affiliation: Faculty of Applied Sciences, No. 1, Jalan Menara Gading, UCSI Heights, Cheras 56000 Kuala Lumpur, Malaysia.
authors.forename: R L H
authors.initials: RL
authors.lastname: Lim
mesh_terms: D000072283:A549 Cells; D000363:Agaricales / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D002449:Cell Aggregation / Q000502:physiology; D045744:Cell Line, Tumor; D004912:Erythrocytes / Q000187:drug effects*; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology*; D006367:HeLa Cells; D006384:Hemagglutination / Q000187:drug effects*; D006801:Humans; D007155:Immunologic Factors / Q000235:genetics / Q000378:metabolism*; D061986:MCF-7 Cells; D051379:Mice; D062105:Molecular Docking Simulation; D017510:Protein Folding
chemical_list: D005656:Fungal Proteins; D007155:Immunologic Factors; C061148:LZ-8 protein, Ganoderma lucidum