title: Engineering of chicken avidin: a progressive series of reduced charge mutants.
abstract: Avidin, a positively charged egg-white glycoprotein, is a widely used tool in biotechnological applications because of its ability to bind biotin strongly. The high pI of avidin (approximately 10.5), however, is a hindrance in certain applications due to non-specific (charge-related) binding. Here we report a construction of a series of avidin charge mutants with pIs ranging from 9.4 to 4.7. Rational design of the avidin mutants was based on known crystallographic data together with comparative sequence alignment of avidin, streptavidin and a set of avidin-related genes which occur in the chicken genome. All charge mutants retained the ability to bind biotin tightly according to optical biosensor interaction analysis. In most cases, their thermal stability characteristics were indistinguishable from those of the wild-type avidin. Our results demonstrate that the charge properties of avidin can be modified without disturbing the crucial biotin-binding activity.
authors.affiliation: Department of Biological and Environmental Science, University of Jyväskylä, Finland.
authors.forename: A T
authors.initials: AT
authors.lastname: Marttila
authors.forename: K J
authors.initials: KJ
authors.lastname: Airenne
authors.forename: O H
authors.initials: OH
authors.lastname: Laitinen
authors.forename: T
authors.initials: T
authors.lastname: Kulik
authors.forename: E A
authors.initials: EA
authors.lastname: Bayer
authors.forename: M
authors.initials: M
authors.lastname: Wilchek
authors.forename: M S
authors.initials: MS
authors.lastname: Kulomaa
mesh_terms: D000818:Animals; D001360:Avidin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D002645:Chickens; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D018411:Spodoptera
chemical_list: D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D001360:Avidin; D001710:Biotin