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ISOLATION, CHARACTERIZATION AND RADIOIMMUNOASSAY Of RAT ALPHA-MACROfETOPROTEIN (ACUTE PHASE-A2 MACROGLOBULIN)

MOLECULAR IMMUNOLOGY 1979年第8期16卷 547-554页

作者： HUDIG, D SELL, S UNIV CALIF SAN DIEGO DEPT PATHOL LA JOLLA CA 92093 USA

Rat alpha-macrofetoprotein (AMf)§ has been isolated and purified by antibody affinity chromatography and quantitated using a double antibody radioimmunoassay (RIA) with a sensitivity of 20 ng. Two forms of AMf, differing in reference mobilities ( R ) in 5% polyacrylamide gels and in isoelectric focusing points (pI), are present in serum. The R f 0.10 AMf has a pI of 4.35 and R f 0.25 AMf a pI of 4.65. R f 0.25 AMf competes 2.2 times better than R f 0.10 AMf in an RIA with 125 I R f 0.25 AMf as the predominant antigen. The two forms do not differ substantially in mol. wt. AMf has a mol. wt of 700,000 ± 70,000 determined by SDS polyacrylamide gel electrophoresis. Dithiothreitol reduction of AMf resulted in six different chains, the largest with mol. wt 175,000. It is suggested that the five smaller chains are the result of interaction of AMf with endoproteases. The E 1% 280 of AMf is 9.16 ± 0.3 by the biuret method using bovine serum albumin as a standard and 10.6 ± 0.3 by microKjeldahl N analysis. Amino-acid analysis of AMf is compared with the analyses recently reported by others. Normal adult rat serum AMf concentrations determined for two groups of rats are 17 ± 2 and 32 ± 6 μg/ml.

关键词： AMf, alpha-macrofetoprotein AAAMf antibody affinity purified macrofetoprotein DEAE diethylaminoethyl cellulose E 1% 280 absorbance at 280 nm of a 1% wt/volume solution IgG immunoglobulin G IgM immunoglobulin M HA 2 M human alpha 2 -macroglobulin NAM normal alpha-macroglobulin pI isoelectric point s.e.m. standard error of the mean R f reference mobility RIA radioimmunoassay SDS sodium dodecyl sulfate

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ANALYSIS Of ELECTROPHORETIC BEHAVIOR Of CYTOCHROME-C OXIDASE SUBUNITS - RETARDATION COEffICIENT VERSUS MOLECULAR-WEIGHT IN DODECYL-SULfATE UREA GELS

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第1期576卷 109-117页

作者： TRACY, RP CHAN, SHP SYRACUSE UNIV DEPT BIOL BIOL RES LABS SYRACUSE NY 13210 USA

Standard proteins were examined by electrophoresis in highly cross-linked polyacrylamide gels containing sodium dodecyl sulfate and urea. Their behavior was analyzed at a single gel concentration (by MW vs. relative mobility) and at several gel concentrations (MW vs. retardation coefficient). The validity of the latter method of analysis was established for this gel system. Beef heart cytochrome c oxidase [EC 1.9.3.1] was subjected to analysis by this method. Compared to standards, subunits I and III showed increased free electrophoretic mobility, while that of subunit V was slightly decreased. The MW values derived were: I, 44,600;II, 22,700;III, 23,500;IV 16,900;V, 9400;VI, 7600;VII, 4300. The SE were all less than .+-. 7%. Isolated V and VI were analyzed by 2 dimensional dodecyl sulfate electrophoresis, in which the 2nd dimension also contained urea. In contrast to their behavior when the holo-enzyme was examined, these isolated subunits V and VI no longer exchange migrating positions relative to each other during the 2 dimensional analysis. The MW values of the isolated subunits agree with those of the holo-enzyme.

关键词： Cytochrome c oxidase subunit Subunit molecular weight Retardation coefficient R f relative mobility f r retardation coefficient M o free mobility (extrapolated Y intercept of the ferguson plot)

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CHEMICAL-REACTIVITY Of METALLOPROTEINS IN CONfORMATIONALLY OUT-Of-EQUILIBRIUM STATES

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第3-4期549卷 255-280页

作者： BLUMENfELD, LA DAVIDOV, RM Institute of Chemical Physics Academy of Science of the U.S.S.R. Moscow U.S.S.R.

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HISTIDYL AND TYROSYL RESIDUE IONIZATION STUDIES Of SUBTILISIN NOVO

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第2期578卷 261-268页

作者： OMAR, S BROWN, Mf SILVER, P SCHLEICH, T UNIV CALIF SANTA CRUZ DIV NAT SCI SANTA CRUZ CA 95064 USA

The low field portion of the 360 MHz 1H NMR spectrum of phenylmethanesulfonyl-subtilisin Novo (EC 3.4.21.14) was studied as a function of pH. Analysis of the pH-induced chemcial shift changes occurring between 6-7 ppm revealed 5 classes of ionizable residues with pK values (uncorrected) of 10.3, 10.7, 10.7, 10.8 and 11.0. With a single exception, the titration curves can be fit by assuming a simple proton ionization equilibrium. four classes of low intensity broad resonances, assigned to the histidyl residues, are observed between 8 and 9 ppm. Uncorrected pK values of 5.4, 5.7, 6.0 and 6.4 were determined for the residues comprising each of these classes. The spectral data are consistent with protonation of one or more histidyl residues upon acid induced denaturation of the protein. These results are compared with those of analogues studies performed by the use of other techniques.

关键词： Subtilisin Novo Serine protease Histidyl residue Tyrosyl residue Titration curve PhCH 2 SO 2 f phenylmethanesulfonyl fluoride PhCh 2 SO 2 - phenylmethanesulfonyl NMR nuclear magnetic resonance ppm parts per million TSP sodium 3-trimethylsilylpropanecarboxylate

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SOME ASPECTS Of ERROR-BOUNDS IN fEATURE-SELECTION

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PATTERN RECOGNITION 1979年第5-6期11卷 353-360页

作者： BOEKEE, DE VANDERLUBBE, JCA Information Theory Group Department of Electrical Engineering Delft University of Technology 4 Mekelweg Delft The Netherlands

In this paper we discuss various bounds on the Bayesian probability of error, which are used for feature selection, and are based on distance measures and information measures. We show that they are basically of two types. One type can be related to the f -divergence, the other can be related to information measures. This also clarifies some properties of these measures for the two-class problem and for the multiclass problem. We give some general bounds on the Bayesian probability of error and discuss various aspects of the different approaches.

关键词： Information measures Distance measures feature evaluation f -divergence Error bounds

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ENERGY-CONVERSION IN THE fUNCTIONAL MEMBRANE Of PHOTOSYNTHESIS - ANALYSIS BY LIGHT-PULSE AND ELECTRIC PULSE METHODS - CENTRAL ROLE Of THE ELECTRIC-fIELD

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第3-4期505卷 355-427页

作者： WITT, HT Max-Volmer-Institut für Physikalische Chemie und Molekularbiologie Technische Universität Berlin Strasse des 17. Juni 135 1 Berlin 12 Germany

The pulse methods are applied to study of the physicochemical foundations of photosynthesis of green plants. Some of the topics in this review are: photoreactions of the chlorophylls;electron transfer;proton transloca... 详细信息

关键词： ΔA field-indicating absorption change and electrochromic shift BChl bacteriochlorophyll Car carotenoid CCCP carbonylcyanide m -chloropheylhydrazone Cf 1 coupling factor of the ATPase Ch1 chlorophyll Chl- a chlorophyll a Chl- b chlorophyll b Chl- a I ( P -700) chlorophyl a I (reaction center of antennae system I) Chl- a II ( P -680) chlorophyl a II (reaction center of antennae system II) Cyte cytochrome DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea DCIP 2,6-dichlorophenoll indophenol Δφ electrical potential difference Δψ surface potential difference f electric filed strength G free energy i ion current i H + H + current i K + K + current j current density PC plastocyanine PMS N -methylphenazonium sulfate PQ (1) plastoquinone primary electron-acceptro of Chl- a II (≡ X -320) PQ (2) plastoquinone, connector between PQ (1) and PQ pool PQ plastoquinone, member of the PQ pool τ half-life and half-rise time X -320 plastoquinone primary electron-acceptro of Chl- a II (≡PQ (1) 1 (…) single turnover conditions ss (…) steady state in saturating permanent light S enzyme system for the cleavage of H 2 O

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IMMUNOLOGICAL AND RECONSTITUTION STUDIES ON THE ADENOSINE-TRIPHOSPHATASE COMPLEX fROM RHODOSPIRILLUM-RUBRUM

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第1期545卷 77-85页

作者： MULLER, HW SCHMITT, M SCHNEIDER, E DOSE, K UNIV MAINZ INST BIOCHEMD-6500 MAINZFED REP GER

Studies on restoration of membrane-bound adenosinetriphosphatase (ATP phosphohydrolase, EC 3.6.1.3) from Rhodospirillum rubrum show that the δ-subunit is capable of binding to the f 1 factor or to the f 0 moiety of the f 0 -f 1 ATPase complex. This subunit is thus likely involved in linking the f 0 and f 1 factor. During solubilization of the oligomycin-sensitive f 0 -f 1 ATPase complex with Triton X-100 the detergent becomes specifically associated with the lipophilic f 0 part of the enzyme complex. Crossed immunoelectrophoresis, agglutination tests, and kinetic studies with anti-f 1 ATPase antibodies reveal a reaction of immunological identity of membrane-bound ATPase, f 0 -f 1 ATPase, and f 1 ATPase.

关键词： ATPase complex f 1 -ATPase Reconstitution (Rhodospirillum rubrum)

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TERNARY COMPLEX-fORMATION Of PIG-HEART LACTATE-DEHYDROGENASE WITH SPIN-LABELED COENZYMES AND INHIBITORS AS STUDIED BY ELECTRON-SPIN RESONANCE

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第2期568卷 287-296页

作者： WENZEL, HR TROMMER, WE RUHR UNIV BOCHUM LEHRSTUHL BIOCHEM CHEM ABT D-4630 BOCHUM FED REP GER

The formation of ternary inhibitor and dead end complexes of pig heart lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) was studied by means of 2 NAD derivatives, spin-labeled at N6 and C-8 of the adenine ring. Dissociation constants calculated for the inhibitors oxamate and oxalate from their corresponding ternary complexes are in excellent agreement with data from literature derived from sedimentation experiments. However, the recently postulated enzyme-NADH-sulfite complex was not observed. The mobility of the spin-label, i.e., the protein conformation near the adenine binding pocket in various ternary complexes, depends on the type of inhibition or substrate employed.

关键词： Lactate dehydrogenase Spin label Ternary complex ESR NAD N 6 SL- or 8SL-preceding NAD denote a nitroxyl spin-label attached to the N 6 - or 8-position of the adenine moiety as shown in fig. 2, SL signifies N 6 SL or 8SL. The subscripts t , b, f with concentration data mean total bound and free respectively

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INACTIVE TO ACTIVE TRANSITIONS Of THE MITOCHONDRIAL ATPASE COMPLEX AS CONTROLLED BY THE ATPASE INHIBITOR

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BIOCHIMICA ET BIOPHYSICA ACTA 1979年第2期547卷 252-257页

作者： GOMEZPUYOU, A DEGOMEZPUYOU, MT ERNSTER, L UNIV STOCKHOLM DEPT BIOCHEM ARRHENIUS LAB S-10691 STOCKHOLM SWEDEN

The hydrolytic and phosphorylation activities of the ATPase complex of bovine heart mitochondria are regulated by the ATPase inhibitor of Pullman and Monroy. The inhibiting action of the peptide on ATPase activity was overcome by a proton-motive force. Submitochondrial particles that contain the inhibitor, either intrinsically or externally added, show a lag that precedes phosphorylation. Particles devoid of the inhibitor or particles that are in an active state fail to present the lag. Prior to the onset of phosphorylation, the ATPase complex probably undergoes a transition to an active state through a process that involves the inhibitor. The transition depends on the concentration of ATP, 50 .mu.M ATP giving 50% inhibition of the proton-motive force-induced transition.

关键词： ATPase ATPase inhibitor Mitochondria Proton-motive force Submitochondrial particle f 1 -ATPase soluble mitochondrial ATPase fCCP carbonylcyanide p -trifluoromethoxy-phenylhydrazone MES 2-( N -morpholino)ethanesulfonic acid

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EffECTS Of PHOSPHOLIPID ACYL CHAIN STRUCTURE ON PHYSICAL-PROPERTIES .1. ISOBRANCHED PHOSPHATIDYLCHOLINES

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CHEMISTRY AND PHYSICS Of LIPIDS 1979年第3期24卷 287-296页

作者： SILVIUS, JR MCELHANEY, RN Department of Biochemistry University of Alberta Edmonton Alberta T6G 2H7 Canada

All isobranched fatty acids of 12-18 C were synthesized in gram quantities by a convenient acetylene coupling reaction followed by catalytic hydrogenation. The corresponding phosphatidylcholines (PC) were synthesized and their thermotropic phase behavior investigated by differential thermal analysis. The isobranched acyl phosphatidylcholines show gel-to-liquid-crystalline phase transition temperatures (Tcs) some ***. C below those of the corresponding straight-chain PC and appear to exhibit 2 slowly interconverting low-temperature phases below Tc. The observed strong alternation of Tc between isobranched PC with odd- and even-C number acyl chains contrasts with the behavior of the straight-chain PC and suggests that the acyl chains of the branched-chain PC are strongly tilted with respect to the bilayer normal below and/or above Tc while those of the straight-chain PC are not. Significant differences apparently exist in the overall packing of branched- and straight-chain PC in the gel and possibly the liquid-crystalline state.

关键词： DMSO dimethyl sulfoxide DTA differential thermal analysis GLC gas-liquid chromatography PC phosphatidylcholine T c temperatures of major heating endotherm for a PC T f temperatures of major cooling exotherm for a PC TLC thin-layer chromatography T m fatty acid melting temperature

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