Characterization of human DAAO variants potentially related to an increased risk of schizophrenia.


Abstract

Considering the key role of d-serine in N-methyl-d-aspartate receptor-mediated neurotransmission, it is highly relevant to define the role that enzymes play in d-serine synthesis and degradation. In particular, the details of regulation of the d-serine catabolic human enzyme d-amino acid oxidase (hDAAO) are unknown although different lines of evidence have shown it to be involved in schizophrenia susceptibility. Here we investigated the effect of three single nucleotide polymorphisms and known mutations in hDAAO, i.e., D31H, R279A, and G331V. A very low amount of soluble G331V hDAAO is produced in E. coli cells: the recombinant variant enzyme is fully active. Human U87 glioblastoma cells transiently transfected for G331V hDAAO show a low viability, a significant amount of protein aggregates, and augmented apoptosis. The recombinant D31H and R279A hDAAO variants do not show alterations in tertiary and quaternary structures, thermal stability, binding affinity for inhibitors, and the modulator pLG72, whereas the kinetic efficiency and the affinity for d-serine and for FAD were higher than for the wild-type enzyme. While these effects for the substitution at position 31 cannot be structurally explained, the R279A mutation might affect the hDAAO FAD-binding affinity by altering the "structurally ambivalent" peptide V47-L51. In agreement with the observed increased activity, expression of D31H and R279A hDAAO variants in U87 cells produces a higher decrease in cellular d/(d+l) serine ratio than the wild-type counterpart. In vivo, these substitutions could affect cellular d-serine concentration and its release at synapsis and thus might be relevant for schizophrenia susceptibility.

Submission Details

ID: qdUmBSr64

Submitter: Shu-Ching Ou

Submission Date: Dec. 3, 2018, 9:41 a.m.

Version: 1

Publication Details
Caldinelli L;Sacchi S;Molla G;Nardini M;Pollegioni L,Biochim Biophys Acta (2013) Characterization of human DAAO variants potentially related to an increased risk of schizophrenia. PMID:23219954
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
3ZNN 2013-05-15 1.9 IN VITRO AND IN VIVO INHIBITION OF HUMAN D-AMINO ACID OXIDASE: REGULATION OF D-SERINE CONCENTRATION IN THE BRAIN
3G3E 2009-06-23 2.2 Crystal structure of human D-amino acid oxidase in complex with hydroxyquinolin-2(1H)
3ZNO 2013-05-15 2.3 IN VITRO AND IN VIVO INHIBITION OF HUMAN D-AMINO ACID OXIDASE: REGULATION OF D-SERINE CONCENTRATION IN THE BRAIN
4QFC 2014-07-16 2.4 Co-crystal structure of compound 3 (4-hydroxy-6-[2-(7-hydroxy-2-oxo-4-phenyl-2h-chromen-6-yl)ethyl]pyridazin-3(2h)-one) and FAD bound to human DAAO at 2.4A
3ZNP 2013-05-15 2.4 IN VITRO AND IN VIVO INHIBITION OF HUMAN D-AMINO ACID OXIDASE: REGULATION OF D-SERINE CONCENTRATION IN THE BRAIN
3CUK 2008-07-22 2.49 Crystal structure of human D-amino acid oxidase: bound to an inhibitor
3W4I 2013-05-29 2.5 Crystal Structure of human DAAO in complex with coumpound 8
2DU8 2006-11-21 2.5 Crystal structure of human D-amino acid oxidase
5ZJ9 2018-10-10 2.6 human D-amino acid oxidase complexed with 5-chlorothiophene-3-carboxylic acid
5ZJA 2018-10-10 2.6 human D-amino acid oxidase complexed with 5-chlorothiophene-2-carboxylic acid
2E4A 2007-03-06 2.6 Crystal Structure of Human D-Amino Acid Oxidase in complex with o-aminobenzoate
2E82 2007-03-06 2.7 Crystal structure of human D-amino acid oxidase complexed with imino-DOPA
3W4J 2013-05-29 2.74 Crystal Structure of human DAAO in complex with coumpound 12
3ZNQ 2013-05-15 2.75 IN VITRO AND IN VIVO INHIBITION OF HUMAN D-AMINO ACID OXIDASE: REGULATION OF D-SERINE CONCENTRATION IN THE BRAIN
4QFD 2014-07-16 2.85 Co-crystal structure of compound 2 (3-(7-hydroxy-2-oxo-4-phenyl-2H-chromen-6-yl)propanoic acid) and FAD bound to human DAAO at 2.85A
3W4K 2013-05-29 2.86 Crystal Structure of human DAAO in complex with coumpound 13
2E48 2007-03-06 2.9 Crystal Structure of Human D-Amino Acid Oxidase: Substrate-Free Holoenzyme
2E49 2007-03-06 3.2 Crystal Structure of Human D-Amino Acid Oxidase in Complex with Imino-Serine

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
375.6 A,B,C,D D-amino-acid oxidase A2V9Y8 OXDA_MACFA
400.0 A,B,C,D D-amino-acid oxidase P14920 OXDA_HUMAN