scholarly journals Structure and substrate specificity determinants of the taurine biosynthetic enzyme cysteine sulphinic acid decarboxylase

2020 ◽  
Author(s):  
Elaheh Mahootchi ◽  
Arne Raasakka ◽  
Weisha Luan ◽  
Gopinath Muruganandam ◽  
Remy Loris ◽  
...  
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Active Site ◽  
Ph Regulation ◽  
Enzymatic Oxidation ◽  
Acid Decarboxylase ◽  
Critical Determinant ◽  
Signalling Molecules ◽  
Binding Cavity ◽  
Abundant Amino Acid

AbstractPyridoxal 5′-phosphate (PLP) is an important cofactor for amino acid decarboxylases with many biological functions, including the synthesis of signalling molecules, such as serotonin, dopamine, histamine, γ-aminobutyric acid, and taurine. Taurine is an abundant amino acid with multiple physiological functions, including osmoregulation, pH regulation, antioxidative protection, and neuromodulation. In mammalian tissues, taurine is mainly produced by decarboxylation of cysteine sulphinic acid to hypotaurine, catalysed by the PLP-dependent cysteine sulphinic acid decarboxylase (CSAD), followed by non-enzymatic oxidation of the product to taurine. We determined the crystal structure of mouse CSAD and compared it to other PLP-dependent decarboxylases in order to identify determinants of substrate specificity and catalytic activity. Recognition of the substrate involves distinct side chains forming the substrate-binding cavity. In addition, the backbone conformation of a buried active-site loop appears to be a critical determinant for substrate side chain binding in PLP-dependent decarboxylases. Phe94 was predicted to affect substrate specificity, and its mutation to serine altered both the catalytic properties of CSAD and its stability. Using small-angle X-ray scattering, we further showed that similarly to its closest homologue, GADL1, CSAD presents open/close motions in solution. The structure of apo-CSAD indicates that the active site gets more ordered upon internal aldimine formation. Taken together, the results highlight details of substrate recognition in PLP-dependent decarboxylases and provide starting points for structure-based inhibitor design with the aim of affecting the biosynthesis of taurine and other abundant amino acid metabolites.

Engineering the Substrate Specificity of Porcine Kidney d-Amino Acid Oxidase by Mutagenesis of the “Active-Site Lid”

2006 ◽  
Vol 139 (5) ◽  
pp. 873-879 ◽  
Author(s):  
Chiaki Setoyama ◽  
Yasuzo Nishina ◽  
Hisashi Mizutani ◽  
Ikuko Miyahara ◽  
Ken Hirotsu ◽  
...  
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Active Site ◽  
Acid Oxidase ◽  
Porcine Kidney ◽  
Amino Acid Oxidase

scholarly journals Structural basis for the histamine synthesis by human histidine decarboxylase

2014 ◽  
Vol 70 (a1) ◽  
pp. C458-C458
Author(s):  
Hirofumi Komori ◽  
Yoko Nitta ◽  
Hiroshi Ueno ◽  
Yoshiki Higuchi
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Histidine Decarboxylase ◽  
The Body ◽  
Structural Basis ◽  
Acid Decarboxylase ◽  
Amino Acid Decarboxylase ◽  
Histamine Production ◽  
Novel Drugs ◽  
High Sequence Homology

Histamine is a bioactive amine responsible for a variety of physiological reactions, including allergy, gastric acid secretion, and neurotransmission. In mammals, histamine production from histidine is catalyzed by histidine decarboxylase (HDC). Mammalian HDC is a pyridoxal 5'-phosphate (PLP)-dependent decarboxylase and belongs to the same family as mammalian glutamate decarboxylase (GAD) and mammalian aromatic L-amino acid decarboxylase (AroDC). The decarboxylases of this family function as homodimers and catalyze the formation of physiologically important amines like GABA and dopamine via decarboxylation of glutamate and DOPA, respectively. Despite high sequence homology, both AroDC and HDC react with different substrates. For example, AroDC catalyzes the decarboxylation of several aromatic L-amino acids, but has little activity on histidine. Although such differences are known, the substrate specificity of HDC has not been extensively studied because of the low levels of HDC in the body and the instability of recombinant HDC, even in a well-purified form. However, knowledge about the substrate specificity and decarboxylation mechanism of HDC is valuable from the viewpoint of drug development, as it could help lead to designing of novel drugs to prevent histamine biosynthesis. We have determined the crystal structure of human HDC in complex with inhibitors, histidine methyl ester (HME) and alpha-fluoromethyl histidine (FMH). These structures showed the detailed features of the PLP-inhibitor adduct (external aldimine) in the active site of HDC. These data provided insight into the molecular basis for substrate recognition among the PLP-dependent L-amino acid decarboxylases.

Substrate specificity of aromatic-L-amino acid decarboxylase

Life Sciences ◽  
1974 ◽  
Vol 14 (5) ◽  
pp. 899-908 ◽  
Author(s):  
Talmage R. Bosin ◽  
Alan R. Buckpitt ◽  
Roger P. Maickel
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Acid Decarboxylase ◽  
Amino Acid Decarboxylase

Gene Cloning, Transcriptional Analysis, Purification, and Characterization of Phenolic Acid Decarboxylase from Bacillus subtilis

1998 ◽  
Vol 64 (4) ◽  
pp. 1466-1471 ◽  
Author(s):  
Jean-François Cavin ◽  
Véronique Dartois ◽  
Charles Diviès
Keyword(s):  
Bacillus Subtilis ◽  
Amino Acid ◽  
Substrate Specificity ◽  
Phenolic Acid ◽  
Genomic Library ◽  
Transcriptional Analysis ◽  
Acid Decarboxylase ◽  
Reading Frame ◽  
Appl Environ ◽  
Phenolic Acid Decarboxylase

ABSTRACT Bacillus subtilis displays a substrate-inducible decarboxylating activity with the following three phenolic acids: ferulic, p-coumaric, and caffeic acids. Based on DNA sequence homologies between the Bacillus pumilus ferulate decarboxylase gene (fdc) (A. Zago, G. Degrassi, and C. V. Bruschi, Appl. Environ. Microbiol. 61:4484–4486, 1995) and theLactobacillus plantarum p-coumarate decarboxylase gene (pdc) (J.-F. Cavin, L. Barthelmebs, and C. Diviès, Appl. Environ. Microbiol. 63:1939–1944, 1997), a DNA probe of about 300 nucleotides for the L. plantarum pdcgene was used to screen a B. subtilis genomic library in order to clone the corresponding gene in this bacterium. One clone was detected with this heterologous probe, and this clone exhibited phenolic acid decarboxylase (PAD) activity. The corresponding 5-kb insertion was partially sequenced and was found to contain a 528-bp open reading frame coding for a 161-amino-acid protein exhibiting 71 and 84% identity with the pdc- and fdc-encoded enzymes, respectively. The PAD gene (pad) is transcriptionally regulated by p-coumaric, ferulic, or caffeic acid; these three acids are the three substrates of PAD. Thepad gene was overexpressed constitutively inEscherichia coli, and the stable purified enzyme was characterized. The difference in substrate specificity between this PAD and other PADs seems to be related to a few differences in the amino acid sequence. Therefore, this novel enzyme should facilitate identification of regions involved in catalysis and substrate specificity.

Investigating the role of active site residues of Rhodotorula gracilis d-amino acid oxidase on its substrate specificity

Biochimie ◽  
2007 ◽  
Vol 89 (3) ◽  
pp. 360-368 ◽  
Author(s):  
Angelo Boselli ◽  
Luciano Piubelli ◽  
Gianluca Molla ◽  
Mirella S. Pilone ◽  
Loredano Pollegioni ◽  
...  
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Active Site ◽  
Acid Oxidase ◽  
Active Site Residues ◽  
Amino Acid Oxidase ◽  
Rhodotorula Gracilis

scholarly journals Amino Acid Residues Contributing to the Substrate Specificity of the Influenza A Virus Neuraminidase

1999 ◽  
Vol 73 (8) ◽  
pp. 6743-6751 ◽  
Author(s):  
Darwyn Kobasa ◽  
Shantha Kodihalli ◽  
Ming Luo ◽  
Maria R. Castrucci ◽  
Isabella Donatelli ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Amino Acid ◽  
Substrate Specificity ◽  
Viral Replication ◽  
Active Site ◽  
Influenza A ◽  
Sialic Acids ◽  
Influenza A Viruses ◽  
Human Viruses

ABSTRACT Influenza A viruses possess two glycoprotein spikes on the virion surface: hemagglutinin (HA), which binds to oligosaccharides containing terminal sialic acid, and neuraminidase (NA), which removes terminal sialic acid from oligosaccharides. Hence, the interplay between these receptor-binding and receptor-destroying functions assumes major importance in viral replication. In contrast to the well-characterized role of HA in host range restriction of influenza viruses, there is only limited information on the role of NA substrate specificity in viral replication among different animal species. We therefore investigated the substrate specificities of NA for linkages betweenN-acetyl sialic acid and galactose (NeuAcα2-3Gal and NeuAcα2-6Gal) and for different molecular species of sialic acids (N-acetyl and N-glycolyl sialic acids) in influenza A viruses isolated from human, avian, and pig hosts. Substrate specificity assays showed that all viruses had similar specificities for NeuAcα2-3Gal, while the activities for NeuAcα2-6Gal ranged from marginal, as represented by avian and early N2 human viruses, to high (although only one-third the activity for NeuAcα2-3Gal), as represented by swine and more recent N2 human viruses. Using site-specific mutagenesis, we identified in the earliest human virus with a detectable increase in NeuAcα2-6Gal specificity a change at position 275 (from isoleucine to valine) that enhanced the specificity for this substrate. Valine at position 275 was maintained in all later human viruses as well as swine viruses. A similar examination of N-glycolylneuraminic acid (NeuGc) specificity showed that avian viruses and most human viruses had low to moderate activity for this substrate, with the exception of most human viruses isolated between 1967 and 1969, whose NeuGc specificity was as high as that of swine viruses. The amino acid at position 431 was found to determine the level of NeuGc specificity of NA: lysine conferred high NeuGc specificity, while proline, glutamine, and glutamic acid were associated with lower NeuGc specificity. Both residues 275 and 431 lie close to the enzymatic active site but are not directly involved in the reaction mechanism. This finding suggests that the adaptation of NA to different substrates occurs by a mechanism of amino acid substitutions that subtly alter the conformation of NA in and around the active site to facilitate the binding of different species of sialic acid.

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