title: Crocodilian tau-crystallin: overexpression, purification, and characterization.
abstract: tau-Crystallin is a taxon-restricted crystallin found in eye lenses of reptiles and a few avian species but presumably absent in mammals. The level of tau-crystallin in the lens varies among different species. In the crocodile lens, it is the least abundant crystallin and is present in trace amounts. We present a method for cloning, overexpression, and purification of crocodilian tau-crystallin utilizing a combination of gel filtration and ion-exchange chromatography yielding an extremely purified protein. The protein gets profusely expressed resulting in a fairly high yield and exists as a monomeric entity of 47.5 kDa molecular mass. The recombinant tau-crystallin exists in a properly folded native state as probed by circular dichroism and fluorescence spectroscopy and exhibits enolase activity.
authors.affiliation: Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, India.
authors.forename: Anurag Kumar
authors.initials: AK
authors.lastname: Mishra
authors.forename: Reena
authors.initials: R
authors.lastname: Chandrashekhar
authors.forename: Ramesh K
authors.initials: RK
authors.lastname: Aggarwal
authors.forename: Yogendra
authors.initials: Y
authors.lastname: Sharma
mesh_terms: D000489:Alligators and Crocodiles; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016622:Silver Staining; D013050:Spectrometry, Fluorescence; D038225:tau-Crystallins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D038225:tau-Crystallins; D010751:Phosphopyruvate Hydratase