Mutation Analysis of the Aspartoacylase Gene in Non-Jewish Patients with Canavan Disease

Zeng, Bai Jin; Pastores, Gregory M.; Leone, Paola; Raghavan, Srinivasa; Wang, Zhaohui; Ribeiro, Lucilene Arilho; Torres, Patrícia; Ong, Elton; Kolodny, Edwin H.

doi:10.1007/0-387-30172-0_11

Cited by 32 publications

(42 citation statements)

References 22 publications

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“…2) depicts the four cysteine residues in ASPA. Mutations at Cys152 have been associated with CD and yield undetectable ASPA activity upon in vitro expression in COS cells (Kaul et al 1996,Zeng et al 2002. Also, low concentrations of DTT are required to maintain ASPA activity, whereas high concentrations ablate activity (Kaul et al 1991,Madhavarao et al 2002.…”

Section: Role Of Cysteines In Aspamentioning

confidence: 99%

“…It has been hypothesized that many non-conservative CD mutants exert deleterious effects due to a loss of structural integrity by a drastic conformational change, rather than directly disrupting catalytic events at the active site (Moore et al 2003,Shaag et al 1995,Zeng et al 2002. Therefore, CD mutations resulting in nearly WT ASPA protein levels likely suggest defective catalysis, while those drastically decreasing ASPA protein suggest something other than defective catalysis.…”

Section: Protein Characteristics Of CD Mutant Aspamentioning

confidence: 99%

“…Notably, missense mutations at His21, Glu24, Asp68, and Arg71 have been detected in CD patients (Sistermans et al 2000,Zeng et al 2002,Janson et al 2006. Although there are no CD mutations at Glu178, nearby CD mutations have been found at Pro181, Pro183, and Val186 (Sistermans et al 2000,Zeng et al 2002,Elpeleg & Shaag 1999, which suggests that Glu178 is an important catalytic residue.…”

Section: Introductionmentioning

confidence: 99%

“…Among Ashkenazi Jews, the E285A mutation is the most common (Elpeleg et al 1994,Feigenbaum et al 2004,Kaul et al 1994,Kaul et al 1996,Kronn et al 1995, and the A305E mutation is the most common one among other populations (Elpeleg & Shaag 1999,Janson et al 2006,Kaul et al 1994,Kaul et al 1996,Shaag et al 1995,Sistermans et al 2000,Yalcinkaya et al 2005,Zeng et al 2002. Less than half of the missense mutations in the ASPA gene have been shown to yield negligible ASPA activity compared with wild-type (WT) enzyme upon transient transfection into COS cells.…”

Section: Introductionmentioning

confidence: 99%

“…These include residues that are equivalent to those involved in zinc coordination (His21, Glu24, His116), shaping the substrate-binding cavity (Asp68), substrate binding (Arg71), transition state stabilization (Arg63), and catalysis (Glu178) (Makarova & Grishin 1999). Notably, missense mutations at His21, Glu24, Asp68, and Arg71 have been detected in CD patients (Sistermans et al 2000,Zeng et al 2002,Janson et al 2006. Although there are no CD mutations at Glu178, nearby CD mutations have been found at Pro181, Pro183, and Val186 (Sistermans et al 2000,Zeng et al 2002,Elpeleg & Shaag 1999, which suggests that Glu178 is an important catalytic residue.…”

Section: Introductionmentioning

confidence: 99%

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Mutational analysis of aspartoacylase: Implications for Canavan Disease

et al. 2007

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Mutations that result in undetectable activity of aspartoacylase, which catalyzes the deacetylation of N-acetyl-L-aspartate, correlate with Canavan Disease, a neurodegenerative disorder usually fatal during childhood. The underlying biochemical mechanisms of how these mutations ablate activity are poorly understood. Therefore, we developed and tested a three-dimensional homology model of aspartoacylase based on zinc dependent carboxypeptidase A. Mutations of the putative zinc-binding residues (H21G, E24D/G, and H116G), the general proton donor (E178A), and mutants designed to switch the order of the zinc-binding residues (H21E/E24H and E24H/H116E) yielded wild-type aspartoacylase protein levels and undetectable ASPA activity. Mutations that affect substrate carboxyl binding (R71N) and transition state stabilization (R63N) also yielded wild-type aspartoacylase protein levels and undetectable aspartoacylase activity. Alanine substitutions of Cys124 and Cys152, residues indicated by homology modeling to be in close proximity and in the proper orientation for disulfide bonding, yielded reduced ASPA protein and activity levels. Finally, expression of several previously tested (E24G, D68A, C152W, E214X, D249V, E285A, and A305E) and untested (H21P, A57T, I143T, P183H, M195R, K213E/G274R, G274R, and F295S) Canavan Disease mutations resulted in undetectable enzyme activity, and only E285A and P183H showed wild-type aspartoacylase protein levels. These results show that aspartoacylase is a member of the caboxypeptidase A family and offer novel explanations for most loss-of-function aspartoacylase mutations associated with Canavan Disease.

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Section: Role Of Cysteines In Aspamentioning

confidence: 99%

Section: Protein Characteristics Of CD Mutant Aspamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mutational analysis of aspartoacylase: Implications for Canavan Disease

et al. 2007

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2‐Chlorodeoxyadenosine and cytarabine combination therapy for idiopathic hypereosinophilic syndrome

Jabbour

Verstovšek

Giles

et al. 2005

Cancer

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BACKGROUNDHypereosinophilic syndrome (HES) is a rare, disabling, and incurable disease. In this study, a combination of 2‐chlorodeoxyadenosine (2‐CdA) and cytosine arabinoside (ara‐C) chemotherapy was evaluated in patients with HES.METHODSNine patients with HES were treated with ara‐C (1 g/m2) given intravenously over 2 hours at 0 hours, 48 hours, 72 hours, 96 hours, and 120 hours; and 2‐CdA (12 mg/m2 per day) was given as a continuous intravenous infusion over 5 days starting at 24 hours. A second course of the same therapy was administered in patients who had a response.RESULTSAll patients had signs and symptoms of end‐organ involvement. The median time from diagnosis to therapy was 25 months. Seven patients had received prior therapies. Five patients (55%) achieved a complete remission (CR), 1 after receiving 2 courses of therapy. Elimination of eosinophilia was accompanied by the resolution of symptoms. The median disease‐free survival and overall survival after the diagnosis for patients who achieved CR was 26 months and 44 months, respectively. Treatment was tolerated well. Febrile neutropenia occurred in 28% of the 14 courses that were given. The median time to recovery from neutropenia and thrombocytopenia was 17 days and 39 days, respectively.CONCLUSIONSThe combined 2‐CdA and ara‐C chemotherapy regimen had activity in patients with HES. Cancer 2005. © 2005 American Cancer Society.

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Clinically Distinct Phenotypes of Canavan Disease Correlate with Residual Aspartoacylase Enzyme Activity

et al. 2017

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We describe 14 patients with 12 novel missense mutations in ASPA, the gene causing Canavan disease (CD). We developed a method to study the effect of these 12 variants on the function of aspartoacylase—the hydrolysis of N‐acetyl‐l‐aspartic acid (NAA) to aspartate and acetate. The wild‐type ASPA open reading frame (ORF) and the ORFs containing each of the variants were transfected into HEK293 cells. Enzyme activity was determined by incubating cell lysates with NAA and measuring the released aspartic acid by LC–MS/MS. Clinical data were obtained for 11 patients by means of questionnaires. Four patients presented with a non‐typical clinical picture or with the milder form of CD, whereas seven presented with severe CD. The mutations found in the mild patients corresponded to the variants with the highest residual enzyme activities, suggesting that this assay can help evaluate unknown variants found in patients with atypical presentation. We have detected a correlation between clinical presentation, enzyme activity, and genotype for CD.

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Mutation Analysis of the Aspartoacylase Gene in Non-Jewish Patients with Canavan Disease

Cited by 32 publications

References 22 publications

Mutational analysis of aspartoacylase: Implications for Canavan Disease

Mutational analysis of aspartoacylase: Implications for Canavan Disease

2‐Chlorodeoxyadenosine and cytarabine combination therapy for idiopathic hypereosinophilic syndrome

Clinically Distinct Phenotypes of Canavan Disease Correlate with Residual Aspartoacylase Enzyme Activity

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