title: Directed evolution of lectins by an improved error-prone PCR and ribosome display method.
abstract: Lectins are useful reagents for the structural characterization of glycans. However, currently available lectins have an apparent drawback in their "repertoire," lacking some critical probes, such as those for sulfated glycans. Thus, engineering lectins with novel specificity would be of great practical value. Here, we describe a directed evolution strategy to tailor novel lectins for novel specificity or biological functions. Our strategy uses a reinforced ribosome display-based selection combined with error-prone PCR to isolate mutants with target specificity and an evanescent-field fluorescence-assisted glycoconjugate microarray to rapidly evaluate the specificity of selected mutants. A successful case of screening a lectin, which has acquired an ability to recognize 6-sulfo-galactose-terminated glycans, is described.
authors.affiliation: Institute of Traditional Chinese Medicine and Natural Products, Jinan University, No. 601, Huangpu Avenue West, Guangzhou, 510632, People's Republic of China, hudan77516@gmail.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Hu
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tateno
authors.forename: Jun
authors.initials: J
authors.lastname: Hirabayashi
mesh_terms: D019020:Directed Molecular Evolution / Q000379:methods*; D005690:Galactose / Q000737:chemistry; D006001:Glycoconjugates / Q000378:metabolism; D037102:Lectins / Q000235:genetics* / Q000378:metabolism; D046228:Microarray Analysis; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics*
chemical_list: D006001:Glycoconjugates; D037102:Lectins; D011134:Polysaccharides; D005690:Galactose