Comparison of two metal-dependent pyruvate aldolases related by convergent evolution: substrate specificity, kinetic mechanism, and substrate channeling.

Abstract:

:HpaI and BphI are two pyruvate class II aldolases found in aromatic meta-cleavage degradation pathways that catalyze similar reactions but are not related in sequence. Steady-state kinetic analysis of the aldol addition reactions and product inhibition assays showed that HpaI exhibits a rapid equilibrium random order mechanism while BphI exhibits a compulsory order mechanism, with pyruvate binding first. Both aldolases are able to utilize aldehyde acceptors two to five carbons in length; however, HpaI showed broader specificity and had a preference for aldehydes containing longer linear alkyl chains or C2-OH substitutions. Both enzymes were able to bind 2-keto acids larger than pyruvate, but only HpaI was able to utilize both pyruvate and 2-ketobutanoate as carbonyl donors in the aldol addition reaction. HpaI lacks stereospecific control producing racemic mixtures of 4-hydroxy-2-oxopentanoate (HOPA) from pyruvate and acetaldehyde while BphI synthesizes only (4S)-HOPA. BphI is also able to utilize acetaldehyde produced by the reduction of acetyl-CoA catalyzed by the associated aldehyde dehydrogenase, BphJ. This aldehyde was directly channeled from the dehydrogenase to the aldolase active sites, with an efficiency of 84%. Furthermore, the BphJ reductive deacylation reaction increased 4-fold when BphI was catalyzing the aldol addition reaction. Therefore, the BphI-BphJ enzyme complex exhibits unique bidirectionality in substrate channeling and allosteric activation.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Wang W,Baker P,Seah SY

doi

10.1021/bi100251u

subject

Has Abstract

pub_date

2010-05-04 00:00:00

pages

3774-82

issue

17

eissn

0006-2960

issn

1520-4995

journal_volume

49

pub_type

杂志文章
  • New class of bacterial membrane oxidoreductases.

    abstract::A new class of bacterial multisubunit membrane-bound electron-transfer complexes has been identified based on biochemical and bioinformatic data. It contains subunits homologous to the three-subunit molybdopterin oxidoreductases and four additional subunits, two of which are c-type cytochromes. The complex was purifie...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi047267l

    authors: Yanyushin MF,del Rosario MC,Brune DC,Blankenship RE

    更新日期:2005-08-02 00:00:00

  • Pre-steady-state kinetics of the microtubule-kinesin ATPase.

    abstract::The pre-steady-state kinetics of the microtubule-kinesin ATPase were investigated by chemical-quench flow methods using the Drosophila kinesin motor domain (K401) expressed in Escherichia coli [Gilbert, S. P., & Johnson, K. A. (1993) Biochemistry 32, 4677-4684]. The results define a minimal mechanism: M.K + ATP in equ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00173a044

    authors: Gilbert SP,Johnson KA

    更新日期:1994-02-22 00:00:00

  • Presence of two rhodopsin intermediates responsible for transducin activation.

    abstract::To identify how many rhodopsin intermediates interact with retinal G-protein transducin, the photobleaching process of chicken rhodopsin has been investigated in the presence or absence of transducin by means of time-resolved low-temperature spectroscopy. Singular value decomposition (SVD) analysis of the spectral dat...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi970932o

    authors: Tachibanaki S,Imai H,Mizukami T,Okada T,Imamoto Y,Matsuda T,Fukada Y,Terakita A,Shichida Y

    更新日期:1997-11-18 00:00:00

  • Polyamide nucleic acid targeted to the primer binding site of the HIV-1 RNA genome blocks in vitro HIV-1 reverse transcription.

    abstract::We report here that polyamide nucleic acid (PNA) as well as a polyamide nucleic acid-DNA chimera complementary to the primer binding site of the HIV-1 genome can completely block priming by tRNA3Lys and consequently the in vitro initiation of reverse transcription by HIV-1 RT. Conventional heating and cooling is not r...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi972197m

    authors: Lee R,Kaushik N,Modak MJ,Vinayak R,Pandey VN

    更新日期:1998-01-20 00:00:00

  • His...Asp catalytic dyad of ribonuclease A: structure and function of the wild-type, D121N, and D121A enzymes.

    abstract::The side chains of histidine and aspartate residues form a hydrogen bond in the active sites of many enzymes. In serine proteases, the His...Asp hydrogen bond of the catalytic triad is known to contribute greatly to catalysis, perhaps via the formation of a low-barrier hydrogen bond. In bovine pancreatic ribonuclease ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi972766q

    authors: Schultz LW,Quirk DJ,Raines RT

    更新日期:1998-06-23 00:00:00

  • Side-chain dynamics of two aromatic amino acids in pancreatic phospholipase A2 as studied by deuterium nuclear magnetic resonance.

    abstract::The flexibility of individual amino acid side chains of pancreatic phospholipase A2 in aqueous and micellar solutions was studied with deuterium nuclear magnetic resonance (2H NMR). Bovine pancreatic phospholipase A2 was selectively deuterated at the aromatic ring systems of Trp-3 and Phe-5 and porcine pancreatic phos...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00334a029

    authors: Allegrini PR,van Scharrenburg GJ,Slotboom AJ,de Haas GH,Seelig J

    更新日期:1985-06-18 00:00:00

  • Fluorescence study of conformational flexibility of RNase S-peptide: distance-distribution, end-to-end diffusion, and anisotropy decays.

    abstract::Frequency-domain fluorescence resonance energy transfer and anisotropy measurements were performed to characterize conformational dynamics of an analog of the RNase S-peptide (residues 1-20). Trp was used as a donor by replacing Phe 8, and a dansyl acceptor group was introduced at position 1 or 18. The distance-distri...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00097a009

    authors: Maliwal BP,Lakowicz JR,Kupryszewski G,Rekowski P

    更新日期:1993-11-23 00:00:00

  • Iron binding to horse spleen apoferritin: a vanadyl ENDOR spin probe study.

    abstract::The role of the protein shell in the formation of the hydrous ferric oxide core of ferritin is poorly understood. A VO2+ spin probe study was undertaken to characterize the initial complex of Fe2+ with horse spleen apoferritin (96% L-subunits). A competitive binding study of VO2+ and Fe2+ showed that the two metals co...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00102a012

    authors: Hanna PM,Chasteen ND,Rottman GA,Aisen P

    更新日期:1991-09-24 00:00:00

  • Substrate specificity, metal binding properties, and spectroscopic characterization of the DapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase from Haemophilus influenzae.

    abstract::The catalytic and structural properties of divalent metal ion cofactor binding sites in the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae were investigated. Co(II)-substituted DapE enzyme was 25% more active than the Zn(II)-loaded form of the enzyme. Interestingly, Mn...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi034845+

    authors: Bienvenue DL,Gilner DM,Davis RS,Bennett B,Holz RC

    更新日期:2003-09-16 00:00:00

  • Posttranslational Chemical Mutagenesis: To Reveal the Role of Noncatalytic Cysteine Residues in Pathogenic Bacterial Phosphatases.

    abstract::The field of chemical site-selective modification of proteins has progressed extensively in recent decades to enable protein functionalization for imaging, drug delivery, and functional studies. In this Perspective, we provide detailed insight into an alternative use of site-selective protein chemistry to probe the ro...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b00639

    authors: Bertoldo JB,Terenzi H,Hüttelmaier S,Bernardes GJL

    更新日期:2018-10-30 00:00:00

  • Fluorescent properties of pyrene bound at specific acylation sites of chicken liver fatty acid synthase.

    abstract::The covalent modification of chicken liver fatty acid synthase by 4-(1-pyrenyl)butyryl-CoA (PBA-CoA), a fluorescent analogue of acetyl- and malonyl-CoA, has been studied. The binding isotherms and the kinetics of inactivation suggest 2 mol of PBA-CoA/mol of enzyme is specifically incorporated into the enzyme. Two clas...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00281a032

    authors: Anderson VE,Hammes GG

    更新日期:1983-06-07 00:00:00

  • Substrate-induced inactivation of a crippled beta-glucosidase mutant: identification of the labeled amino acid and mutagenic analysis of its role.

    abstract::The beta-glucosidase from Agrobacterium sp. catalyzes the hydrolysis of beta-glucosides via a covalent alpha-D-glucopyranosyl-enzyme intermediate involving Glu358. Hydrolysis of 2,4-dinitrophenyl beta-D-glucopyranoside by the low activity Glu358Asp mutant of Agrobacterium beta-glucosidase is accompanied by time-depend...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00044a033

    authors: Gebler JC,Trimbur DE,Warren AJ,Aebersold R,Namchuk M,Withers SG

    更新日期:1995-11-07 00:00:00

  • Lysine 2,3-aminomutase: rapid mix-freeze-quench electron paramagnetic resonance studies establishing the kinetic competence of a substrate-based radical intermediate.

    abstract::Lysine 2,3-aminomutase from Clostridia catalyzes the interconversion of L-lysine and L-beta-lysine. The enzyme contains iron-sulfur clusters and is activated by pyridoxal 5'-phosphate and S-adenosylmethionine, all of which participate in catalysis. Current spectroscopic evidence implicates two substrate-based organic ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi960850k

    authors: Chang CH,Ballinger MD,Reed GH,Frey PA

    更新日期:1996-08-27 00:00:00

  • Hematin-hematin self-association states involved in the formation and reactivity of the malaria parasite pigment, hemozoin.

    abstract::The malaria parasite pigment, hemozoin, is a crystal of ferriprotoporphyrin IX (FP-Fe(III)), a product of hemoglobin digestion. Hemozoin formation is essential for FP-Fe(III) detoxification in the parasite; it is the main target of quinoline antimalarials and can modulate immune and inflammation responses. To gain fur...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi100567j

    authors: Klonis N,Dilanian R,Hanssen E,Darmanin C,Streltsov V,Deed S,Quiney H,Tilley L

    更新日期:2010-08-10 00:00:00

  • Protein kinase C acylation by palmitoyl coenzyme A facilitates its translocation to membranes.

    abstract::Protein kinase C (PKC) translocation to specific subcellular membrane loci after cellular stimulation is mediated, in part, through its interaction with diacylglycerol, phosphatidylserine, and calcium. Herein, we present multiple lines of evidence which demonstrate that purified rabbit brain PKC undergoes specific acy...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi980565w

    authors: Ford DA,Horner CC,Gross RW

    更新日期:1998-08-25 00:00:00

  • Exploring the Evolutionary History of Kinetic Stability in the α-Lytic Protease Family.

    abstract::In addition to encoding the tertiary fold and stability, the primary sequence of a protein encodes the folding trajectory and kinetic barriers that determine the speed of folding. How these kinetic barriers are encoded is not well understood. Here, we use evolutionary sequence variation in the α-lytic protease (αLP) p...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.0c00720

    authors: Nixon CF,Lim SA,Sailer ZR,Zheludev IN,Gee CL,Kelch BA,Harms MJ,Marqusee S

    更新日期:2021-01-26 00:00:00

  • Directed evolution of a quorum-quenching lactonase from Mycobacterium avium subsp. paratuberculosis K-10 in the amidohydrolase superfamily.

    abstract::The PLL(PTE-like lactonase)-group of enzymes within the amidohydrolase superfamily hydrolyze N-acyl-homoserine lactones (AHLs) that are involved in bacterial quorum-sensing pathways. These enzymes possess the (beta/alpha)(8)-barrel fold and serve as attractive templates for in vitro evolution and engineering of quorum...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9004045

    authors: Chow JY,Wu L,Yew WS

    更新日期:2009-05-26 00:00:00

  • Spectroscopic Investigation of Cysteamine Dioxygenase.

    abstract::Thiol dioxygenases are mononuclear non-heme FeII-dependent metalloenzymes that initiate the oxidative catabolism of thiol-containing substrates to their respective sulfinates. Cysteine dioxygenase (CDO), the best characterized mammalian thiol dioxygenase, contains a three-histidine (3-His) coordination environment rat...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.0c00267

    authors: Fernandez RL,Dillon SL,Stipanuk MH,Fox BG,Brunold TC

    更新日期:2020-07-07 00:00:00

  • Partitioning of fish and insect antifreeze proteins into ice suggests they bind with comparable affinity.

    abstract::Antifreeze proteins (AFPs) inhibit the growth of ice by binding to the surface of ice crystals, preventing the addition of water molecules to cause a local depression of the freezing point. AFPs from insects are much more effective at depressing the freezing point than fish AFPs. Here, we have investigated the possibi...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi035605x

    authors: Marshall CB,Tomczak MM,Gauthier SY,Kuiper MJ,Lankin C,Walker VK,Davies PL

    更新日期:2004-01-13 00:00:00

  • Structure around the cleavage site in the thrombin receptor determined by NMR spectroscopy.

    abstract::NMR spectroscopic experiments were performed to study the structure of synthetic peptides identical to two extracellular regions of the human thrombin receptor. The smaller molecule, comprising 14 amino acids, was biologically active and was equivalent to the "tethered ligand" exposed after cleavage of the receptor by...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00186a005

    authors: Smith KJ,Trayer IP,Grand RJ

    更新日期:1994-05-24 00:00:00

  • Conformational properties of B-Z junctions in DNA.

    abstract::The structural consequences of specific base sequences in DNA can exert a strong influence on the function of DNA. It has previously been reported that the presence of multiple B-Z conformational junctions in constructed DNA oligomers results in unusually enhanced electrophoretic gel mobilities of these oligomers [Win...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00134a026

    authors: Lu M,Guo Q,Kallenbach NR,Sheardy RD

    更新日期:1992-05-19 00:00:00

  • Effects of guanidine hydrochloride on the proton inventory of proteins: implications on interpretations of protein stability.

    abstract::The DeltaG degrees (N)(-)(D) value obtained from extrapolation to zero denaturant concentration by the linear extrapolation method (LEM) is commonly interpreted to represent the Gibbs energy difference between native (N) and denatured (D) ensembles at the limit of zero denaturant concentration. For DeltaG degrees (N)(...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi001071d

    authors: Bolen DW,Yang M

    更新日期:2000-12-12 00:00:00

  • NMR characterization of three-disulfide variants of lysozyme, C64A/C80A, C76A/C94A, and C30A/C115A--a marginally stable state in folded proteins.

    abstract::Our earlier NMR study showed that a two-disulfide variant of hen lysozyme containing intra-alpha-domain disulfide bridges, C6-C127 and C30-C115, is partially folded, with the alpha domain tightly folded to the nativelike conformation and the beta domain flexible or unfolded. With a view that the formation of a third d...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi049967w

    authors: Yokota A,Hirai K,Miyauchi H,Iimura S,Noda Y,Inoue K,Akasaka K,Tachibana H,Segawa S

    更新日期:2004-06-01 00:00:00

  • Structural Changes of the Active Center during the Photoactivation of Xenopus (6-4) Photolyase.

    abstract::Photolyases (PHRs) repair the UV-induced photoproducts, cyclobutane pyrimidine dimer (CPD) or pyrimidine-pyrimidone (6-4) photoproduct [(6-4) PP], restoring normal bases to maintain genetic integrity. CPD and (6-4) PP are repaired by substrate-specific PHRs, CPD PHR and (6-4) PHR, respectively. Flavin adenine dinucleo...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b01111

    authors: Yamada D,Yamamoto J,Zhang Y,Iwata T,Hitomi K,Getzoff ED,Iwai S,Kandori H

    更新日期:2016-02-02 00:00:00

  • Correlating structure and affinity for PEX5:PTS1 complexes.

    abstract::Many proteins that are destined to reside within the lumen of the peroxisome contain the peroxisomal targeting signal-1 (PTS1), a C-terminal tripeptide approximating the consensus sequence -Ser-Lys-Leu-COO(-). The PTS1 is recognized by the tetratricopeptide repeat (TPR) domains of PEX5, a cytosolic receptor that cycle...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi027034z

    authors: Gatto GJ Jr,Maynard EL,Guerrerio AL,Geisbrecht BV,Gould SJ,Berg JM

    更新日期:2003-02-18 00:00:00

  • Separation and quantitation of intracellular forms of poliovirus RNA by agarose gel electrophoresis.

    abstract::Intracellular poliovirus-specific RNA species can be measured directly by electrophoresis of total cytoplasmic nucleic acids through 1% agarose gels, resulting in the separation of single- and double-stranded forms of poliovirus RNA from each other and from HeLa cell 28S ribosomal RNA. Single-stranded RNA molecules di...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00631a027

    authors: Hewlett MJ,Rozenblatt S,Ambros V,Baltimore D

    更新日期:1977-06-14 00:00:00

  • Structural Analysis of Cj1427, an Essential NAD-Dependent Dehydrogenase for the Biosynthesis of the Heptose Residues in the Capsular Polysaccharides of Campylobacter jejuni.

    abstract::Many strains of Campylobacter jejuni display modified heptose residues in their capsular polysaccharides (CPS). The precursor heptose was previously shown to be GDP-d-glycero-α-d-manno-heptose, from which a variety of modifications of the sugar moiety have been observed. These modifications include the generation of 6...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.0c00096

    authors: Huddleston JP,Anderson TK,Spencer KD,Thoden JB,Raushel FM,Holden HM

    更新日期:2020-04-07 00:00:00

  • Demonstration of nitrogen coordination in metal--bleomycin complexes by electron spin--echo envelope spectroscopy.

    abstract::We have studied the Cu(II), Co(II), and Fe(III) complexes of the antineoplastic drug bleomycin by using electron spin--echo envelope spectroscopy. For all three complexes, nitrogen coordination of the metal ions is demonstrated. For the Cu(II)-- and Co(II)--drug complexes, we have been able to identify imidazole as a ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00509a045

    authors: Burger RM,Adler AD,Horwitz SB,Mims WB,Peisach J

    更新日期:1981-03-17 00:00:00

  • Structure and dynamics of an acid-denatured protein G mutant.

    abstract::NMR studies of protein denatured states provide insights into potential initiation sites for folding that may be too transient to be observed kinetically. We have characterized the structure and dynamics of the acid-denatured state of protein G by using a F30H mutant of G(B1) which is on the margin of stability. At 5 ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9920230

    authors: Sari N,Alexander P,Bryan PN,Orban J

    更新日期:2000-02-08 00:00:00

  • Exchange mechanisms for hydrogen bonding protons of cytidylic and guanylic acids.

    abstract::The pH dependence of buffer catalysis of exchange of the C-4 amino protons of cyclic cytosine 2',3'-monophosphate (cCMP) and the N-1 proton of cyclic guanosine 2',3'-monophosphate (cGMP) conforms to an exchange mechanism, in which protonation of the nucleobases at C(N-3) AND G(N-7) establishes the important intermedia...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00608a035

    authors: McConnell B

    更新日期:1978-07-25 00:00:00