Publications

• Genome-wide analysis of substrate specificities of the Escherichia coli haloacid dehalogenase-like phosphatase family.

  Kuznetsova E., Proudfoot M., Gonzalez C.F., Brown G., Omelchenko M.V., Borozan I., Carmel L., Wolf Y.I., Mori H., Savchenko A.V., Arrowsmith C.H., Koonin E.V., Edwards A.M., Yakunin A.F.

  Haloacid dehalogenase (HAD)-like hydrolases are a vast superfamily of largely uncharacterized enzymes, with a few members shown to possess phosphatase, beta-phosphoglucomutase, phosphonatase, and dehalogenase activities. Using a representative set of 80 phosphorylated substrates, we characterized ... >> More

  Haloacid dehalogenase (HAD)-like hydrolases are a vast superfamily of largely uncharacterized enzymes, with a few members shown to possess phosphatase, beta-phosphoglucomutase, phosphonatase, and dehalogenase activities. Using a representative set of 80 phosphorylated substrates, we characterized the substrate specificities of 23 soluble HADs encoded in the Escherichia coli genome. We identified small molecule phosphatase activity in 21 HADs and beta-phosphoglucomutase activity in one protein. The E. coli HAD phosphatases show high catalytic efficiency and affinity to a wide range of phosphorylated metabolites that are intermediates of various metabolic reactions. Rather than following the classical "one enzyme-one substrate" model, most of the E. coli HADs show remarkably broad and overlapping substrate spectra. At least 12 reactions catalyzed by HADs currently have no EC numbers assigned in Enzyme Nomenclature. Surprisingly, most HADs hydrolyzed small phosphodonors (acetyl phosphate, carbamoyl phosphate, and phosphoramidate), which also serve as substrates for autophosphorylation of the receiver domains of the two-component signal transduction systems. The physiological relevance of the phosphatase activity with the preferred substrate was validated in vivo for one of the HADs, YniC. Many of the secondary activities of HADs might have no immediate physiological function but could comprise a reservoir for evolution of novel phosphatases. << Less

  J. Biol. Chem. 281:36149-36161(2006) [PubMed] [EuropePMC]

  This publication is cited by 7 other entries.

• A hexose-1-phosphatase in silkworm blood.

  FAULKNER P.

  Biochem J 60:590-596(1955) [PubMed] [EuropePMC]

• Yihx-encoded haloacid dehalogenase-like phosphatase HAD4 from Escherichia coli is a specific alpha-d-glucose 1-phosphate hydrolase useful for substrate-selective sugar phosphate transformations.

  Pfeiffer M., Wildberger P., Nidetzky B.

  Phosphomonoester hydrolases (phosphatases; EC 3.1.3.) often exhibit extremely relaxed substrate specificity which limits their application to substrate-selective biotransformations. In search of a phosphatase catalyst specific for hydrolyzing α-d-glucose 1-phosphate (αGlc 1-<i>P</i>), we selected ... >> More

  Phosphomonoester hydrolases (phosphatases; EC 3.1.3.) often exhibit extremely relaxed substrate specificity which limits their application to substrate-selective biotransformations. In search of a phosphatase catalyst specific for hydrolyzing α-d-glucose 1-phosphate (αGlc 1-<i>P</i>), we selected haloacid dehalogenase-like phosphatase 4 (HAD4) from <i>Escherichia coli</i> and obtained highly active recombinant enzyme through a fusion protein (Z_basic2_HAD4) that contained Z_basic2, a strongly positively charged three α-helical bundle module, at its N-terminus. Highly pure Z_basic2_HAD4 was prepared directly from <i>E. coli</i> cell extract using capture and polishing combined in a single step of cation exchange chromatography. Kinetic studies showed Z_basic2_HAD4 to exhibit 565-fold preference for hydrolyzing αGlc 1-<i>P</i> (<i>k</i>_cat/<i>K</i>_M = 1.87 ± 0.03 mM^-1 s^-1; 37 °C, pH 7.0) as compared to d-glucose 6-phosphate (Glc 6-<i>P</i>). Also among other sugar phosphates, αGlc 1-<i>P</i> was clearly preferred. Using different mixtures of αGlc 1-<i>P</i> and Glc 6-<i>P</i> (e.g. 180 mM each) as the substrate, Z_basic2_HAD4 could be used to selectively convert the αGlc 1-<i>P</i> present, leaving back all of the Glc 6-<i>P</i> for recovery. Z_basic2_HAD4 was immobilized conveniently using direct loading of <i>E. coli</i> cell extract on sulfonic acid group-containing porous carriers, yielding a recyclable heterogeneous biocatalyst that was nearly as effective as the soluble enzyme, probably because protein attachment to the anionic surface occurred in a preferred orientation via the cationic Z_basic2 module. Selective removal of αGlc 1-<i>P</i> from sugar phosphate preparations could be an interesting application of Z_basic2_HAD4 for which readily available broad-spectrum phosphatases are unsuitable. << Less

  J. Mol. Catal., B Enzym. 110:39-46(2014) [PubMed] [EuropePMC]

• The hydrolysis of glucose monophosphates by a phosphatase preparation from pea seeds.

  TURNER D.H., TURNER J.F.

  Biochem J 74:486-491(1960) [PubMed] [EuropePMC]