title: α-Crystallin protects human arginosuccinate lyase activity under freeze-thaw conditions.
abstract: Argininosuccinate lyase (ASL) catalyzes the conversion of argininosuccinate into arginine and fumarate, a key step in the biosynthesis of urea and arginine. ASL is a tetrameric enzyme but it dissociates into inactive dimers under low temperature conditions. This study investigates the inactivation process under low temperature conditions. Inactivation was caused by dissociation of tetrameric ASL into dimers, with increased exposure of hydrophobic areas without disturbance of the secondary structure or the microenvironment surrounding the key tryptophan residues. Most activity was retained when temperatures were changed at a rate of >1 °C/min, whilst freezing or thawing more slowly resulted in greater loss of activity. Inactivation was reduced by inclusion of α-crystallin, a structural protein found in ocular lenses and a member of the small heat-shock protein family, by stabilization of the ASL quaternary structure. In addition, α-crystallin was able to restore the function of ASL that had been inactivated by slow freezing and thawing. The effect of α-crystallin was similar to that of bovine serum albumin, suggesting that both proteins exerted their effects by hydrophobic interactions. α-Crystallin therefore acts as a cryo-preservative that protects ASL activity during freezing and thawing.
authors.affiliation: Department of Biochemistry, National Defense Medical Center, No. 161, Sec. 6, MinChuan E. Rd., Neihu, Taipei, Taiwan.
authors.forename: Yi-Yu
authors.initials: YY
authors.lastname: Wei
authors.forename: Chih-Wei
authors.initials: CW
authors.lastname: Huang
authors.forename: Wei-Yuan
authors.initials: WY
authors.lastname: Chou
authors.forename: Hwei-Jen
authors.initials: HJ
authors.lastname: Lee
mesh_terms: D000818:Animals; D001123:Argininosuccinate Lyase / Q000737:chemistry*; D002942:Circular Dichroism; D003080:Cold Temperature; D003451:Cryoprotective Agents / Q000737:chemistry*; D004926:Escherichia coli; D005615:Freezing; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013552:Swine; D038201:alpha-Crystallins / Q000737:chemistry*
chemical_list: D003451:Cryoprotective Agents; D011994:Recombinant Proteins; D012996:Solutions; D038201:alpha-Crystallins; D012710:Serum Albumin, Bovine; D001123:Argininosuccinate Lyase