SASDGS4: Human alpha-aminoadipic semialdehyde dehydrogenase (ALDH)7A1 E399...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDGS4

試料

Human alpha-aminoadipic semialdehyde dehydrogenase (ALDH)7A1 E399G at 1.6 mg/mL

• Alpha-aminoadipic semialdehyde dehydrogenase E399G (protein), ALDH7A1 E399G, Homo sapiens

機能・相同性

分子機能:

L-aminoadipate-semialdehyde dehydrogenase / L-aminoadipate-semialdehyde dehydrogenase activity / Choline catabolism / choline catabolic process / Lysine catabolism / betaine-aldehyde dehydrogenase / betaine-aldehyde dehydrogenase (NAD+) activity / glycine betaine biosynthetic process from choline / aldehyde metabolic process / glyceraldehyde-3-phosphate dehydrogenase (NAD+) (non-phosphorylating) activity / aldehyde dehydrogenase (NAD+) / aldehyde dehydrogenase (NAD+) activity / sensory perception of sound / mitochondrial matrix / mitochondrion / extracellular exosome / identical protein binding / nucleus / cytosol

ドメイン・相同性:

Aldehyde dehydrogenase family 7 member A1-like / Aldehyde dehydrogenase, glutamic acid active site / Aldehyde dehydrogenases glutamic acid active site. / Aldehyde dehydrogenase domain / Aldehyde dehydrogenase family / Aldehyde dehydrogenase, C-terminal / Aldehyde dehydrogenase, N-terminal / Aldehyde/histidinol dehydrogenase

構成要素:

Alpha-aminoadipic semialdehyde dehydrogenase

• 生物種: Homo sapiens (ヒト)
• 引用: ジャーナル: J Inherit Metab Dis / 年: 2020; タイトル: Structural analysis of pathogenic mutations targeting Glu427 of ALDH7A1, the hot spot residue of pyridoxine-dependent epilepsy.; 著者: Adrian R Laciak / David A Korasick / Kent S Gates / John J Tanner / ; 要旨: Certain loss-of-function mutations in the gene encoding the lysine catabolic enzyme aldehyde dehydrogenase 7A1 (ALDH7A1) cause pyridoxine-dependent epilepsy (PDE). Missense mutations of Glu427, especially Glu427Gln, account for ~30% of the mutated alleles in PDE patients, and thus Glu427 has been referred to as a mutation hot spot of PDE. Glu427 is invariant in the ALDH superfamily and forms ionic hydrogen bonds with the nicotinamide ribose of the NAD cofactor. Here we report the first crystal structures of ALDH7A1 containing pathogenic mutations targeting Glu427. The mutant enzymes E427Q, Glu427Asp, and Glu427Gly were expressed in Escherichia coli and purified. The recombinant enzymes displayed negligible catalytic activity compared to the wild-type enzyme. The crystal structures of the mutant enzymes complexed with NAD were determined to understand how the mutations impact NAD binding. In the E427Q and E427G structures, the nicotinamide mononucleotide is highly flexible and lacks a defined binding pose. In E427D, the bound NAD adopts a "retracted" conformation in which the nicotinamide ring is too far from the catalytic Cys residue for hydride transfer. Thus, the structures revealed a shared mechanism for loss of function: none of the variants are able to stabilise the nicotinamide of NAD in the pose required for catalysis. We also show that these mutations reduce the amount of active tetrameric ALDH7A1 at the concentration of NAD tested. Altogether, our results provide the three-dimensional molecular structural basis of the most common pathogenic variants of PDE and implicate strong (ionic) hydrogen bonds in the aetiology of a human disease.

登録者

• David Korasick (Mizzou, University of Missouri-Columbia, Columbia, MO, USA)

モデル

• モデル #3627: ; タイプ: atomic / カイ2乗値: 0.738301313217999; Omokage検索でこの集合体の類似形状データを探す (詳細)
• モデル #3628: ; タイプ: atomic / カイ2乗値: 0.738301313217999; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: Human alpha-aminoadipic semialdehyde dehydrogenase (ALDH)7A1 E399G at 1.6 mg/mL; 試料濃度: 1.4 mg/ml
• バッファ: 名称: 50 mM HEPES, 100 mM NaCl, 1 mM DTT, 10 mM NAD, 5% (v/v) glycerol; pH: 8

要素 #1848

名称: ALDH7A1 E399G / タイプ: protein / 記述: Alpha-aminoadipic semialdehyde dehydrogenase E399G / 分子量: 55.488 / 由来: Homo sapiens / 参照: UniProt: P49419
配列:

GHMSTLLINQ PQYAWLKELG LREENEGVYN GSWGGRGEVI TTYCPANNEP IARVRQASVA DYEETVKKAR EAWKIWADIP APKRGEIVRQ IGDALREKIQ VLGSLVSLEM GKILVEGVGE VQEYVDICDY AVGLSRMIGG PILPSERSGH ALIEQWNPVG LVGIITAFNF PVAVYGWNNA IAMICGNVCL WKGAPTTSLI SVAVTKIIAK VLEDNKLPGA ICSLTCGGAD IGTAMAKDER VNLLSFTGST QVGKQVGLMV QERFGRSLLE LGGNNAIIAF EDADLSLVVP SALFAAVGTA GQRCTTARRL FIHESIHDEV VNRLKKAYAQ IRVGNPWDPN VLYGPLHTKQ AVSMFLGAVE EAKKEGGTVV YGGKVMDRPG NYVEPTIVTG LGHDASIAHT GTFAPILYVF KFKNEEEVFA WNNEVKQGLS SSIFTKDLGR IFRWLGPKGS DCGIVNVNIP TSGAEIGGAF GGEKHTGGGR ESGSDAWKQY MRRSTCTINY SKDLPLAQGI KFQ

実験情報

• ビーム: 設備名称: Advanced Light Source (ALS) 12.3.1 (SIBYLS) / 地域: Berkeley, CA / 国: USA / 線源: X-ray synchrotron / 波長: 0.127 Å / スペクトロメータ・検出器間距離: 2 mm
• 検出器: 名称: Pilatus3 X 2M / Pixsize x: 172 mm

スキャン

タイトル: Human alpha-aminoadipic semialdehyde dehydrogenase (ALDH)7A1 E399G at 1.6 mg/mL
測定日: 2019年5月1日 / 保管温度: 4 °C / セル温度: 10 °C / 単位: 1/A /

	Min	Max
Q	0.0109	0.3938

結果

カーブのタイプ: single_conc /

	実験値	Porod
分子量	196 kDa	-
体積	-	290 nm3

	P(R)	Guinier	Guinier error
前方散乱 I0	22.67	22.83	0.09
慣性半径, Rg	3.72 nm	3.78 nm	0.02

	Min	Max
D	-	11.3
Guinier point	1	43