Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.5.1.12 (biotinidase)
 392 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Folding of DNA into chromatin is mediated by binding to histones such as H4; association of DNA with histones is regulated by covalent histone modifications, e.g. acetylation, methylation, and biotinylation. We sought to identify amino-acid residues that are biotinylated in histone H4, and to determine whether acetylation and methylation of histones affect biotinylation. Synthetic peptides spanning fragments of human histone H4 were biotinylated enzymatically using biotinidase. Peptide-bound biotin was probed with streptavidin-peroxidase. Peptides based on the N-terminal sequence of histone H4 were effectively recognized by biotinidase as substrates for biotinylation; in contrast, peptides based on the C-terminal sequences were not biotinylated. Substitution of K8 or K12 with alanine or arginine decreased biotinylation, suggesting that these lysines are targets for biotinylation; K8 and K12 are also known targets for acetylation. Chemical acetylation or methylation of a given lysine decreased subsequent enzymatic biotinylation of neighboring lysines, consistent with cross-talk among histone modifications. Substitution of a given lysine (positive charge) with glutamate (negative charge) abolished biotinylation of neighboring lysines, providing evidence that the net charge of histones has a role in biotinylation. An antibody was generated that specifically recognized histone H4 biotinylated at K12. This antibody was used to detect biotinylated histone H4 in nuclear extracts from human cells. These studies suggest that K8 and K12 in histone H4 are targets for biotinylation, that acetylation and biotinylation compete for the same binding sites, and that acetylation and methylation of histones affect biotinylation of neighboring lysines.
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PMID:K8 and K12 are biotinylated in human histone H4. 1515 16

Untreated individuals with deficient activity of biotinidase, the enzyme responsible for recycling the vitamin biotin, usually exhibit neurological and cutaneous findings. To better understand the variability in expression of the disorder it is important to understand the structure of the enzyme and the putative effects of various mutations on its activity. Past attempts to express and purify sufficient quantities of the enzyme by us and others have failed. Therefore, we have resorted to computer modeling using homologous related, crystallized nitrilases/amidases to predict the 3-dimensional structure of biotinidase. The resultant structure is a two domain protein with the catalytic triad consisting of glutamate, lysine and cysteine, within the larger domain. The model predicts multiple glycosylation sites at the surface of the enzyme and multiple disulfide bonds. The precise location of the biotin-binding site could not be determined. Characteristics of 45 missense mutations known to cause profound and partial biotinidase deficiency were examined, including their location, their distance from the catalytic triad, and their potential effect on the structure of the enzyme. Although there are obviously short-comings in predicting the 3-dimensional structure of a protein without crystallographic data, because of the marked homology between biotinidase and specific crystallized amidases/nitrilases, the predicted 3-dimensional structure of biotinidase is probable and should be useful providing clues to structure-function relationships and ultimately the effect of mutations on altering the enzyme's hydrolase and transferase activities.
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PMID:Three dimensional structure of human biotinidase: computer modeling and functional correlations. 1762 31