Thermostabilization of ovalbumin by alkaline treatment: Examination of the possible roles of D-serine residues.


Abstract

It was revealed from the crystal structure analysis of S-ovalbumin (S-OVA) formed by alkaline treatment that Ser164, Ser236, and Ser320 take the D-amino acid residue configuration (Yamasaki et al., J Biol Chem 2003; 278:35524-35530). To address the implications of a D-configuration for these Ser residues in S-OVA formation, three mutant OVAs (S164A, S236A, and S320A) were generated to compare their thermostabilities before and after alkaline treatment. Following alkaline treatment, S236A showed a marked increase in melting temperature similar to the wild type (DeltaT(m), +9 degrees C) which corresponded to the formation of S-OVA, whereas the increment in T(m) for both S164A and S320A was only 4.5 degrees C. Furthermore, the T(m) value of the double mutant S164/320A remained unchanged after alkaline treatment, supporting the relevance of Ser164 and Ser320 for thermostabilization of OVA. As Arg142 was predicted to interact with D-Ser164 upon S-OVA formation, it was substituted to Ala to generate R142A. The resulting increment in T(m) of mutant R142A after alkaline treatment was 5.8 degrees C. The double mutant R142/S320A was therefore prepared to eliminate the participation of Ser320 in thermostabilization, and its T(m) value was compared before and after alkaline treatment. As expected, the increase in T(m) for the double mutant was only 1.2 degrees C. Taken together, the data suggest that D-configuration of Ser164 caused by alkaline treatment favors interaction with Arg142 through conformational changes of the side chain. These results strongly supported the participation of the configurational inversion of both Ser164 and Ser320 residues in the formation of S-OVA. Study holds ProTherm entries: 25617, 25618, 25619, 25620, 25621, 25622, 25623, 25624, 25625, 25626, 25627, 25628, 25629, 25630 Extra Details: ovalbumin; S-ovalbumin; D-amino acid configuration; alkaline treatment

Submission Details

ID: ZYEZ5W3z3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:55 p.m.

Version: 1

Publication Details
Ishimaru T;Ito K;Tanaka M;Matsudomi N,Protein Sci. (2010) Thermostabilization of ovalbumin by alkaline treatment: Examination of the possible roles of D-serine residues. PMID:20512973
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1UHG 2003-07-22 1.9 Crystal Structure of S-Ovalbumin At 1.9 Angstrom Resolution
1OVA 1992-04-15 1.95 CRYSTAL STRUCTURE OF UNCLEAVED OVALBUMIN AT 1.95 ANGSTROMS RESOLUTION
3P9M 2011-10-19 2.0 Crystal Structure of H2-Kb in complex with a mutant of the chicken ovalbumin epitope OVA-G4
3P9L 2011-10-19 2.0 Crystal Structure of H2-Kb in complex with the chicken ovalbumin epitope OVA
3PAB 2011-10-19 2.2 Crystal Structure of H2-Kb in complex with a mutant of the chicken ovalbumin epitope OVA-E1
1JTI 2001-09-05 2.3 Loop-inserted Structure of P1-P1' Cleaved Ovalbumin Mutant R339T
1VAC 1996-06-20 2.5 MHC CLASS I H-2KB HEAVY CHAIN COMPLEXED WITH BETA-2 MICROGLOBULIN AND CHICKEN OVALBUMIN
3CVH 2008-09-23 2.9 How TCR-like antibody recognizes MHC-bound peptide
1P4L 2003-11-11 2.9 Crystal structure of NK receptor Ly49C mutant with its MHC class I ligand H-2Kb
3C8K 2008-04-15 2.9 The crystal structure of Ly49C bound to H-2Kb
4HKJ 2012-11-14 3.0 Structure of Cowpox CPXV203 in complex with MHCI (H-2Kb)
1P1Z 2003-11-11 3.26 X-RAY CRYSTAL STRUCTURE OF THE LECTIN-LIKE NATURAL KILLER CELL RECEPTOR LY-49C BOUND TO ITS MHC CLASS I LIGAND H-2Kb

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.7 Ovalbumin O73860 OVAL_MELGA
100.0 Ovalbumin P01012 OVAL_CHICK