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Biochemical and Structural Analysis of Substrate Specificity of a Phenylalanine Ammonia-Lyase

PLANT PHYSIOLOGY 2018年第2期176卷 1452-1468页

作者： Jun, Se-Young Sattler, Steven A. Cortez, Gabriel S. Vermerris, Wilfred Sattler, Scott E. Kanga, ChulHee Washington State Univ Dept Chem Pullman WA 99164 USA Washington State Univ Sch Mol Biosci Pullman WA 99163 USA Univ Florida Inst Food & Agr Sci Dept Microbiol & Cell Sci Gainesville FL 32610 USA Univ Florida Genet Inst Gainesville FL 32610 USA Univ Nebraska USDA ARS Wheat Sorghum & Forage Res Unit Lincoln NE 68583 USA Univ Nebraska Dept Agron & Hort Lincoln NE 68583 USA

Phenylalanine ammonia-lyase (PAL) is the first enzyme of the general phenylpropanoid pathway catalyzing the nonoxidative elimination of ammonia from L-phenylalanine to give trans-cinnamate. In monocots, PAL also displays tyrosine ammonia lyase (TAL) activity, leading to the formation of p-coumaric acid. The catalytic mechanism and substrate specificity of a major PAL from sorghum (Sorghum bicolor;SbPAL1), a strategic plant for bioenergy production, were deduced from crystal structures, molecular docking, site-directed mutagenesis, and kinetic and thermodynamic analyses. This first crystal structure of a monocotyledonous PAL displayed a unique conformation in its flexible inner loop of the 4-methylidene-imidazole-5-one (MIO) domain compared with that of dicotyledonous plants. The side chain of histidine-123 in the MIO domain dictated the distance between the catalytic MIO prosthetic group created from (189)Ala-Ser-Gly(191) residues and the bound L-phenylalanine and L-tyrosine, conferring the deamination reaction through either the Friedel-Crafts or E-2 reaction mechanism. Several recombinant mutant SbPAL1 enzymes were generated via structure-guided mutagenesis, one of which, H123F-SbPAL1, has 6.2 times greater PAL activity without significant TAL activity. Additional PAL isozymes of sorghum were characterized and categorized into three groups. Taken together, this approach identified critical residues and explained substrate preferences among PAL isozymes in sorghum and other monocots, which can serve as the basis for the engineering of plants with enhanced biomass conversion properties, disease resistance, or nutritional quality.

关键词： SHCBP1 gene TAL Substrate Specificity Phenylalanine Phenylalanine Ammonia-Lyase FRIEDEL CRAFTS Sorghum Liliopsida programmable logic arrays Biochemicals Substrate preferences LRIT1 gene crystal structure

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Synthesis of 3-deoxyanthocyanidin phytoalexins in sorghum occurs independent of light

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PHYSIOlogicAL AND MOLECULAR PLANT PATHOLOGY 1996年第6期49卷 377-388页

作者： Weiergang, I Hipskind, JD Nicholson, RL PURDUE UNIV DEPT BOT & PLANT PATHOL W LAFAYETTE IN 47907 USA

Aetiolated mesocotyls of sorghum were inoculated with the anthracnose pathogen, Colletotrichum sublineolum, and placed in the light or in the dark for 120 h. A time course study showed that each of the four known 3-dexoyanthocyanidin phytoalexins accumulated regardless of whether seedlings had been incubated in the light or the dark. Analyses of extracts by high performance liquid chromatography and plasma desorption mass spectrometry confirmed the presence of each of the phytoalexins. The phytoalexins were never detected in the control, uninoculated plants, regardless of the light treatment. Total RNA was extracted from the mesocotyls, and a time course of RNA hybridization revealed accumulation of transcripts for phenylalanine ammonia-lyase (PAL) and chalcone synthase at times consistent with those expected to be required for phytoalexin synthesis. PAL transcripts accumulated to higher levels in light-grown than in dark-grown seedlings. Importantly, chalcone synthase transcripts accumulated in response to inoculation and accumulation was independent of light. The accumulation of phytoalexins in the dark and the increase of RNA transcripts question the assumption that light is always required to initiate phenolic compound synthesis at the level of PAL. The results suggest that sorghum the induction of the synthesis of PAL transcripts and the resultant synthesis of the 3-deoxyanthocyanidin phytoalexins occurs as a response to fungal infection and probably is separate from the induction of PAL and phenolic compound synthesis which occurs as a response to light. (C) 1996 Academic Press Limited.

关键词： plant diseases plant pathogens plant pathogenic fungi defence mechanisms phytoalexins light cereals plant pathology sorghum fungi LRIT1 gene Sorghum Independent Colletotrichum sublineolum SHCBP1 gene Defence mechanisms programmable logic arrays Phytoalexins plant pathogenic fungi Plant pathogen synthesis fungal wilt Plant Diseases

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CMOS-SOS AUTOMATED UNIVERSAL ARRAY

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IEEE JOURNAL OF SOLID-STATE CIRCUITS 1981年第5期16卷 563-570页

作者： BORGINI, F SUSKIND, BA RCA Advanced Technology Laboratories Camden NJ USA

The CMOS/SOS automated universal array (AUA) is a new, minimum-cost approach for generating custom LSI devices. This is achieved by implementing a compatible automatic placement and routing program which automatically generates a single, customized interconnect metal level that defines the desired logical function. The AUA topological design is specifically configured to be compatible with automatic layout techniques, and as a result the AUA system can guarantee 100 percent connectivity with greater than 90 percent gate utilization for any random-logic application. The CMOS/SOS AUA is an equivalent 800-gate design consisting of a repetitive pattern of 640 basic internal cells and 62 basic peripheral cells for I/O functions, as well as a variety of special cells located in the periphery which meet the needs of most digital applications that may be encountered. Depending on the logic design complexity and computer used, an artwork tape depicting the metallized layout pattern can be generated within 24 h.

关键词： logic arrays Routing logic design programmable logic arrays Application software Automatic logic units Large scale integration logic devices Layout logic functions

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FUNCTIONALLY PARALLEL ARCHITECTURE FOR ARRAY PROCESSORS

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COMPUTER 1981年第9期14卷 28-&页

作者： COHLER, EU STORER, JE CSPI

Based on the natural division of mathematical problems, functional parallelism becomes the architectural key for improving speed/cost ratios for array processors.

关键词： Parallel architectures Hardware Parallel processing programmable logic arrays Concurrent computing Parallel programming Floating-point arithmetic logic arrays Clustering algorithms Functional programming

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HIGH-SPEED ESFI SOS programmable logic ARRAY WITH AN MNOS VERSION

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IEEE JOURNAL OF SOLID-STATE CIRCUITS 1975年第5期10卷 331-336页

作者： HORNINGER, K SIEMENS AG RES LABSMUNICHFED REP GER

A programmable logic array (PLA) with J-K flip-flops as feedback loops and having a maximum operating speed of 12 MHz has been designed and realized in epitaxial-silicon-films-on-insulators (ESFI) silicon-on-sapphire (SOS) technology. The advantages of the ESFI SOS technology and the circuit of the PLA are described and experimental results are presented. In addition, a twin PLA using metal-nitride-oxide-semiconductor (MNOS) transistors in the AND and OR matrices and having the same number of inputs, outputs, and feedback loops as the mask-programmable PLA has been designed. This MNOS PLA has full on-chip decoding capability and can be programmed or reprogrammed individually. The circuit of the MNOS PLA is described and the speed of the device is calculated.

关键词： programmable logic arrays Read only memory logic design Equations logic circuits Feedback loop Decoding Production logic programming Integrated circuit technology

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AN ELECTRICALLY ALTERABLE PLA FOR FAST TURNAROUND-TIME VLSI DEVELOPMENT HARDWARE

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IEEE JOURNAL OF SOLID-STATE CIRCUITS 1981年第5期16卷 570-577页

作者： WOOD, RA HSIEH, YN PRICE, CA WANG, PP System Communications Division IBM Kingston NY USA IBM Corporate Headquarters Armonk NY USA

An electrically alterable PLA (EAPLA) has been designed in double polysilicon FET technology to serve as fast turnaround-time development hardware for custom VLSI designs. Included are descriptions of the cell design, the unique circuitry that is used, the chip architecture and a description of the PLA-based design system incorporating the EAPLA. The PLA architecture consists of two arrays cascaded to perform AND-OR combinational logic functions. It has 24 inputs (2-bit partitioning), 76 product terms, and 28 outputs. Sixteen of the product terms and all of the outputs are `dottable' off-chip with modules of the same type to meet larger PLA requirements. Four modules can be dotted at each array output.

关键词： programmable logic arrays Very large scale integration Hardware Circuits System testing Nonvolatile memory logic functions Design automation logic design Computer architecture

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Anatomically determined functional conduction delay in the posterior left atrium: Relationship to structural heart disease

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JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY 2008年第8期51卷 856-862页

作者： Roberts-Thomson, Kurt C. Stevenson, Irene H. Kistler, Peter M. Haqqani, Haris M. Goldblatt, John C. Sanders, Prashanthan Kalman, Jonathan M. Royal Melbourne Hosp Dept Cardiol Melbourne Vic 3050 Australia Royal Melbourne Hosp Dept Cardiothorac Surg Melbourne Vic 3050 Australia Univ Melbourne Melbourne Vic Australia Royal Adelaide Hosp Dept Cardiol Cardiovasc Res Ctr Adelaide SA 5000 Australia Univ Adelaide Discipline Med Adelaide SA Australia

Objectives This study sought to characterize the conduction properties of the posterior left atrium (PLA) in patients with different forms of structural heart disease undergoing cardiac surgery. Background The PLA plays an important role in the initiation and maintenance of atrial fibrillation. Methods This study included 34 patients having elective cardiac surgery. There were 4 groups of patients: normal left ventricular (LV) function (coronary artery bypass grafting [CABG]);severe LV dysfunction (LVF/CABG);severe mitral regurgitation (MR);severe aortic stenosis (AS). Epicardial mapping of the PLA was performed in sinus rhythm and during differential pacing. Activation patterns, regional conduction velocity (CV), conduction heterogeneity, anisotropy, and total plaque activation time (TAT) were assessed. Results Left atrial size in patients with LVF/CABG (47 +/- 7 mm) and MR (54 +/- 6 mm) was larger than patients with CABG (39 +/- 7 mm) and AS (42 +/- 6 mm;p < 0.05). During pacing, all patients developed a vertical line of conduction delay running between the pulmonary veins. The extent of this conduction delay was greater in patients with LVF/CABG and MR than patients with AS and CABG (p < 0.05). Conduction heterogeneity, anisotropy, and TAT were greater in patients with LVF/CABG and MR than patients with CABG (p < 0.05). These changes resulted in circuitous wave front propagation. Conclusions There is a line of functional conduction delay in a consistent anatomical location in the PLA in patients with structural heart disease. This is most marked in conditions associated with significant chronic atrial enlargement and leads to circuitous wave front propagation, suggesting a potential role in arrhythmogenesis.

关键词： Aged Atrial Fibrillation Coronary Artery Disease Electrophysiology Female 老年人心房颤动电生理学心脏传导系统心脏瓣膜疾病时间因素 Left Atrium tat Genes Epicardial Mapping Atrial Fibrillation Cardiovascular Diseases Electrophysiology programmable logic arrays Heart Diseases interatrial block Coronary Artery Disease atrial fibrillation Intra-Atrial Conduction Delay circuity Patients

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REORGANIZING CIRCUITS TO AID TESTABILITY

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IEEE DESIGN & TEST OF COMPUTERS 1991年第3期8卷 49-57页

作者： GUPTA, R SRINIVASAN, R BREUER, MA UNIV SO CALIF DEPT ELECT ENGN SYSTLOS ANGELESCA 90089

A method of partitioning a circuit canonically into disjoint subcircuits and to group similarly connected storage elements is described. The method is being used in a program called Crete (for Clouding, hierarchical Reorganization, Equivalence determination, Test-methodology embedding, and Editing). Crete partitions and reorganizes the hierarchical description of a circuit so that the designer can apply design-for-testability or built-in self-test techniques to the new hierarchy. The partitioning preserves the designer's circuit hierarchy as much as possible to allow the easy identification of equivalence among partitioned groups. The method reduces the time needed for both test generation and test application in designs that use full scan, partial scan, and built-in self-test techniques. Results for a Viterbi decoder validate the concepts underlying Crete.

关键词： Circuit testing Clouds System testing logic testing Registers programmable logic arrays Built-in self-test logic design Time division multiplexing Hardware

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COOPERATION - JAPAN NEW WATCHWORD

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IEEE MICRO 1993年第5期13卷 90-92页

作者： KAHANER, DK U.S. Office of Naval Research USA

As Japan attempts to keep pace with developments in the changing world of high technology and computer science, its government and industry leaders are moving on several fronts simultaneously. Efforts underway in several industries to improve coordination and cooperation between the public and private sector, as well as with foreign entities, are described. The industries include the ASIC chip, digital compact cassette and minidisc, construction, ISDN, and computer industries.< >

关键词： Computer industry programmable logic arrays Research and development Government Educational institutions Application specific integrated circuits Field programmable gate arrays Protection logic devices Economic indicators

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PHYSIOlogicAL-ACTIVITY OF 1-AMINO-2-PHENYLETHYLPHOSPHONIC ACID, A SUBSTRATE-ANALOG OF PHENYLALANINE

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BIOLOGIA PLANTARUM 1986年第2期28卷 91-+页

作者： KNYPL, JS JANAS, KM 1.Plant Growth Substances Laboratory University of Lod? ul. Banacha 12/16 PL-90 237 Lod? Poland

1-Amino-2-phenylethylphosphonic acid (PheP) retards growth in Spirodela oligorrhiza causing morphological malformations, inhibits chlorophyll synthesis in progeny fronds, and markedly stimulates L-phenylalanine ammonia-lyase (PAL) activity in vivo and inhibits it in vitro;in prolonged experiments ribonuclease activity is stimulated. PheP severely inhibits anthrocyanin synthesis in seedlings of red cabbage, with moderate inhibition of PAL activity in vivo;chlorophyll synthesis and growth of the seedlings are little affected.

关键词： SHCBP1 gene Phenylalanine frond Phenylalanine Ammonia-Lyase Analog Analog PHYSIOlogicAL ACTIVITY programmable logic arrays synthesis growth Seedling LRIT1 gene RED CABBAGE Chlorophyll Ribonucleases

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