C-Terminal end and aminoacid Lys48 in HMG-CoA lyase are involved in substrate binding and enzyme activity

Carrasco, P.; Menao, Sebastián; López-Viñas, Eduardo; Santpere, Gabriel; Clotet, Josep; Sierra, Adriana Y.; Gratacós, Esther; Puisac, Beatriz; Gómez‐Puertas, Paulino; Hegardt, Fausto G.; Pié, Juan; Casals, Núria

doi:10.1016/j.ymgme.2007.03.007

Cited by 10 publications

(11 citation statements)

References 25 publications

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“…The fact that all studied missense mutations cause a loss of enzyme activity greater than 95% is consistent with other published data (Mitchell, et al, 1998;Carrasco, et al, 2007) and suggests that the illness appears only in very severe genotypes, and that partial disruption of the enzyme is probably compatible with normal function. Therefore, it is very difficult to establish genotype-phenotype correlations because we only see the effects of very severe genotypes.…”

Section: Genotype-phenotype Correlationssupporting

confidence: 91%

“…These results confirm the Mediterranean mutation as the second most frequent mutation in the world (31 cases), with a specific location in the Iberian Peninsula, where it was carried by 100% of the Portuguese patients (13 cases) (Carrasco, et al, 2007; and by 61% of Spanish (11 cases) (Casale, et al, 1998;Puisac, et al, 2005). This mutation was also found in an Argentinian patient of Spanish ancestry and in one Moroccoan; and since the other studied Moroccoan patient carries it , the Mediterranean mutation may have a high incidence in that country.…”

Section: Mutational Updatesupporting

confidence: 77%

“…In 23 patients mutations were identifyed in both alleles, and in patient 17 only in one allele. Previously, in patient 22, c.144G>T (K48N) mutation of one allele was reported (Carrasco, et al, 2007). In all the patients in which parental DNA was available (cases 6, 7, 8, 9, 10, 11, 14, 15, 17 and 18), inheritance was confirmed.…”

Section: Resultsmentioning

confidence: 75%

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Ten novelHMGCLmutations in 24 patients of different origin with 3-hydroxy-3-methyl-glutaric aciduria

et al. 2009

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3-Hydroxy-3-methylglutaric aciduria is a rare autosomal recessive genetic disorder that affects ketogenesis and L-leucine catabolism. The clinical acute symptoms include vomiting, convulsions, metabolic acidosis, hypoketotic hypoglycaemia and lethargy. To date, 33 mutations in 100 patients have been reported in the HMGCL gene. In this study 10 new mutations in 24 patients are described. They include: 5 missense mutations: c.109G>A, c.425C>T, c.521G>A, c.575T>C and c.598A>T, 2 nonsense mutations: c.242G>A and c.559G>T, one small deletion: c.853delC, and 2 mutations in intron regions: c.497+4A>G and c.750+1G>A. Two prevalent mutations were detected, 109G>T (E37X) in 38% of disease alleles analyzed and c.504_505delCT in 10% of them. Although patients are mainly of European origin (71%) and mostly Spanish (54%), the group is ethnically diverse and includes, for the first time, patients from Pakistan, Palestine and Ecuador. We also present a simple, efficient method to express the enzyme and we analyze the possible functional effects of missense mutations. The finding that all identified missense mutations cause a >95% decrease in the enzyme activity, indicates that the disease appears only in very severe genotypes."

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Section: Genotype-phenotype Correlationssupporting

confidence: 91%

Section: Mutational Updatesupporting

confidence: 77%

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Ten novelHMGCLmutations in 24 patients of different origin with 3-hydroxy-3-methyl-glutaric aciduria

et al. 2009

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“…2B). The C terminus of the lyase molecule lies Ͼ25 Å from the adenine ring of the substrate, contrary to the results obtained from the modeled structure of the enzymeligand complex (25). Instead, the C terminus is interacting with the glycine-rich loop of the other monomer (Fig.…”

Section: Resultsmentioning

confidence: 64%

“…Role of Lys 48 -Lys 48 of HMGCL has been implicated in an inherited K48N mutation (25). In addition, proteomics results have shown that this residue is a site of post-translational acetylation (26).…”

Section: Resultsmentioning

confidence: 99%

Functional Insights into Human HMG-CoA Lyase from Structures of Acyl-CoA-containing Ternary Complexes

Runquist

Montgomery

et al. 2010

Journal of Biological Chemistry

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HMG-CoA lyase (HMGCL 4 ; EC 4.1.3.4) catalyzes a cationdependent cleavage of substrate into acetyl-CoA and acetoacetate (Scheme 1) (1). This reaction is a key step in ketogenesis, the products of which support anaplerotic metabolism in bacteria (2) and energy production in nonhepatic animal tissues (3). Ketogenesis is particularly important to human metabolism during the prenatal period and during fasting or starvation. In accordance with these physiological roles, it is not surprising that gene knock-out in mice results in embryonic lethality (4). The physiological importance of the enzyme in humans is underscored by the observation that mutations that diminish HMGCL activity correlate with inherited metabolic disease that can be lethal if uncontrolled (5).A variety of human mutations, including many point mutations in protein-coding exons of the gene, have been documented (6). A computational modeling approach was used to explain the molecular basis for some mutations linked to inherited disease (7). This led to the prediction that HMGCL adopts a ␤/␣-barrel fold and a proposal that the acyl-S-pantetheine moiety of the bound substrate passes through the barrel lumen. Initial structural work on human HMGCL liganded to cation and hydroxyglutarate (8) demonstrated that the folding prediction was reasonable but that the substrate binding proposal was unlikely to be correct. The positions of bound cation and hydroxyglutarate (from hydrolyzed hydroxyglutaryl-CoA) indicated the catalytic site to be positioned at the C-terminal end of the barrel, but the absence of a full acyl-CoA molecule in the experimentally determined structure limited detailed insight into the substrate-binding site.To more fully address questions regarding the conformation of bound substrate, activator cation liganding, details concerning reaction chemistry and specificity, as well as the molecular basis for certain inherited HMGCL deficiencies, new structural information on enzyme bound to an intact acyl-CoA molecule is required. To remedy this need, complexes of the WT enzyme with the competitive inhibitor 3-hydroxyglutaryl-CoA and also of catalytically deficient R41M enzyme with the authentic substrate HMG-CoA have been supplemented with the activator cation Mg 2ϩ , and crystallization of the desired ternary complexes has been accomplished. Diffraction quality crystals have been produced, supporting three-dimensional structural determinations for ternary complexes of enzyme, cation, and either the acyl-CoA substrate or inhibitory analog. These findings are

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Molecular genetics of HMG-CoA lyase deficiency

Pié

López-Viñas

Puisac

et al. 2007

Molecular Genetics and Metabolism

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C-Terminal end and aminoacid Lys48 in HMG-CoA lyase are involved in substrate binding and enzyme activity

Cited by 10 publications

References 25 publications

Ten novelHMGCLmutations in 24 patients of different origin with 3-hydroxy-3-methyl-glutaric aciduria

Ten novelHMGCLmutations in 24 patients of different origin with 3-hydroxy-3-methyl-glutaric aciduria

Functional Insights into Human HMG-CoA Lyase from Structures of Acyl-CoA-containing Ternary Complexes

Molecular genetics of HMG-CoA lyase deficiency

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