title: Lentiavidins: Novel avidin-like proteins with low isoelectric points from shiitake mushroom (Lentinula edodes).
abstract: A biotin-binding protein with a low isoelectric point (pI), which minimizes electrostatic non-specific binding to substances other than biotin, is potentially valuable. To obtain such a protein, we screened hundreds of mushrooms, and detected strong biotin-binding activity in the fruit bodies of Lentinula edodes, shiitake mushroom. Two cDNAs, each encoding a protein of 152 amino acids, termed lentiavidin 1 and lentiavidin 2 were cloned from L. edodes. The proteins shared sequence identities of 27%-49% with other biotin-binding proteins, and many residues that directly associate with biotin in streptavidin were conserved in lentiavidins. The pI values of lentiavidin 1 and lentiavidin 2 were 3.9 and 4.4, respectively; the former is the lowest pI of the known biotin-binding proteins. Lentiavidin 1 was expressed as a tetrameric protein with a molecular mass of 60 kDa in an insect cell-free expression system and showed biotin-binding activity. Lentiavidin 1, with its pI of 3.9, has a potential for broad applications as a novel biotin-binding protein.
authors.affiliation: Plant Innovation Center, Japan Tobacco, Inc., 700 Higashibara, Iwata, Shizuoka 438-0802, Japan. Electronic address: yoshimitsu.takakura@jt.com.
authors.forename: Yoshimitsu
authors.initials: Y
authors.lastname: Takakura
authors.affiliation: Plant Innovation Center, Japan Tobacco, Inc., 700 Higashibara, Iwata, Shizuoka 438-0802, Japan.
authors.forename: Kozue
authors.initials: K
authors.lastname: Sofuku
authors.affiliation: Plant Innovation Center, Japan Tobacco, Inc., 700 Higashibara, Iwata, Shizuoka 438-0802, Japan.
authors.forename: Masako
authors.initials: M
authors.lastname: Tsunashima
authors.affiliation: Graduate School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kuwata
mesh_terms: D000595:Amino Acid Sequence; D001360:Avidin / Q000737:chemistry* / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D048690:Fruiting Bodies, Fungal / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007526:Isoelectric Point; D008970:Molecular Weight; D020942:Shiitake Mushrooms / Q000737:chemistry* / Q000235:genetics; D055672:Static Electricity; D019809:Streptavidin / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D018076:DNA, Complementary; D005656:Fungal Proteins; C017071:biotin-binding proteins; D001360:Avidin; D001710:Biotin; D019809:Streptavidin
keywords: Biotin-binding protein; Cell-free extract; Isoelectric point; Lentinula edodes; Non-specific binding