Sequential degradation of alphaII and betaII spectrin by calpain in glutamate or maitotoxin-stimulated cells.


:Calpain-catalyzed proteolysis of II-spectrin is a regulated event associated with neuronal long-term potentiation, platelet and leukocyte activation, and other processes. Calpain proteolysis is also linked to apoptotic and nonapoptotic cell death following excessive glutamate exposure, hypoxia, HIV-gp120/160 exposure, or toxic injury. The molecular basis for these divergent consequences of calpain action, and their relationship to spectrin proteolysis, is unclear. Calpain preferentially cleaves II spectrin in vitro in repeat 11 between residues Y1176 and G1177. Unless stimulated by Ca++ and calmodulin (CaM), betaII spectrin proteolysis in vitro is much slower. We identify additional unrecognized sites in spectrin targeted by calpain in vitro and in vivo. Bound CaM induces a second II spectrin cleavage at G1230*S1231. BetaII spectrin is cleaved at four sites. One cleavage only occurs in the absence of CaM at high enzyme-to-substrate ratios near the betaII spectrin COOH-terminus. CaM promotes II spectrin cleavages at Q1440*S1441, S1447*Q1448, and L1482*A1483. These sites are also cleaved in the absence of CaM in recombinant II spectrin fusion peptides, indicating that they are probably shielded in the spectrin heterotetramer and become exposed only after CaM binds alphaII spectrin. Using epitope-specific antibodies prepared to the calpain cleavage sites in both alphaII and betaII spectrin, we find in cultured rat cortical neurons that brief glutamate exposure (a physiologic ligand) rapidly stimulates alphaII spectrin cleavage only at Y1176*G1177, while II spectrin remains intact. In cultured SH-SY5Y cells that lack an NMDA receptor, glutamate is without effect. Conversely, when stimulated by calcium influx (via maitotoxin), there is rapid and sequential cleavage of alphaII and then betaII spectrin, coinciding with the onset of nonapoptotic cell death. These results identify (i) novel calpain target sites in both alphaII and betaII spectrin; (ii) trans-regulation of proteolytic susceptibility between the spectrin subunits in vivo; and (iii) the preferential cleavage of alphaII spectrin vs betaII spectrin when responsive cells are stimulated by engagement of the NMDA receptor. We postulate that calpain proteolysis of spectrin can activate two physiologically distinct responses: one that enhances skeletal plasticity without destroying the spectrin-actin skeleton, characterized by preservation of betaII spectrin; or an alternative response closely correlated with nonapoptotic cell death and characterized by proteolysis of betaII spectrin and complete dissolution of the spectrin skeleton.






Glantz SB,Cianci CD,Iyer R,Pradhan D,Wang KK,Morrow JS




Has Abstract


2007-01-16 00:00:00












  • Mechanistic studies on full length and the catalytic domain of the tandem SH2 domain-containing protein tyrosine phosphatase: analysis of phosphoenzyme levels and Vmax stimulatory effects of glycerol and of a phosphotyrosyl peptide ligand.

    abstract::SHP-1, a protein tyrosine phosphatase containing two tandem SH2 domains, is autoinhibited at rest by its N-terminal SH2 (N-SH2) domain. Relief from autoinhibition and a subsequent 10-60-fold increase in V(max) have been observed upon N-SH2 domain engagement by a specific phosphotyrosyl ligand or upon deletion of the S...


    pub_type: 杂志文章


    authors: Wang J,Walsh CT

    更新日期:1997-03-11 00:00:00

  • Effects of macromolecular crowding on burst phase kinetics of cytochrome c folding.

    abstract::Excluded volume and viscosity effects of crowding agents that mimic crowded conditions in vivo on "classical" burst phase folding kinetics of cytochrome c are assessed in vitro. Upon electron transfer-triggered folding of reduced cytochrome c, far-UV time-resolved circular dichroism (TRCD) is used to monitor folding u...


    pub_type: 杂志文章


    authors: Chen E,Christiansen A,Wang Q,Cheung MS,Kliger DS,Wittung-Stafshede P

    更新日期:2012-12-11 00:00:00

  • Effect of removing the amino-terminal hexapeptide of tropomyosin on the properties of the thin filament.

    abstract::The role of the amino-terminal region of alpha-striated muscle tropomyosin has been analyzed by reconstituting thin filaments with a version of the protein lacking the first six amino acids. While Omp T-digested tropomyosin (residues 7-284) does not bind significantly to F-actin at micromolar concentrations, an intera...


    pub_type: 杂志文章


    authors: Goonasekara CL,Heeley DH

    更新日期:2009-04-21 00:00:00

  • Nitrogenase reactivity: cyanide as substrate and inhibitor.

    abstract::We have examined the reduction of cyanide by using the purified component proteins of nitrogenase (Av1 and Av2). The previously reported self-inhibition phenomenon was found to be an artifact. One of the two species present in cyanide solutions, CN-, was shown to be a potent reversible inhibitor (Ki = 27 microM) of to...


    pub_type: 杂志文章


    authors: Li J,Burgess BK,Corbin JL

    更新日期:1982-08-31 00:00:00

  • Lipid bilayer partitioning and stability of camptothecin drugs.

    abstract::The intense intrinsic fluorescence emissions from several clinically relevant camptothecin drugs have been exploited in order to determine (1) the structural basis of drug binding to lipid bilayers, (2) the lipid bilayer stability of each drug's alpha-hydroxylactone moiety, a pharmacophore which is essential for antit...


    pub_type: 杂志文章


    authors: Burke TG,Mishra AK,Wani MC,Wall ME

    更新日期:1993-05-25 00:00:00

  • Exploring the cellulose/xylan specificity of the beta-1,4-glycanase cex from Cellulomonas fimi through crystallography and mutation.

    abstract::The retaining beta-1,4-glycanase Cex from Cellulomonas fimi, a family 10 glycosyl hydrolase, hydrolyzes xylan 40-fold more efficiently than cellulose. To gain insight into the nature of its preference for xylan, we determined the crystal structure of the Cex catalytic domain (Cex-cd) trapped as its covalent 2-deoxy-2-...


    pub_type: 杂志文章


    authors: Notenboom V,Birsan C,Warren RA,Withers SG,Rose DR

    更新日期:1998-04-07 00:00:00

  • Role of histidine-40 in ribonuclease T1 catalysis: three-dimensionalstructures of the partially active His40Lys mutant.

    abstract::Histidine-40 is known to participate in phosphodiester transesterification catalyzed by the enzyme ribonuclease T1. A mutant enzyme with a lysine replacing the histidine-40 (His40Lys RNase T1) retains considerable catalytic activity [Steyaert, J., Hallenga, K., Wyns, L., & Stanssens, P. (1990) Biochemistry 29, 9064-90...


    pub_type: 杂志文章


    authors: Zegers I,Verhelst P,Choe HW,Steyaert J,Heinemann U,Saenger W,Wyns L

    更新日期:1992-11-24 00:00:00

  • Conformational transition required for efficient splicing of transcripts from hybrid lambda promoter yeast tRNA gene fusion.

    abstract::Fusion of a prokaryotic promoter to a yeast tRNA gene provides a means for uncoupling analyses of mutations affecting splicing from requirements for transcription and other processing steps. For this purpose, a phage lambda promoter was fused to the Saccharomyces cerevisiae tRNATyr(SUP3a) coding sequence. This fusion ...


    pub_type: 杂志文章


    authors: Shapero MH,Greer CL

    更新日期:1991-07-02 00:00:00

  • Structural characterization of lactoperoxidase in the heme environment by proton NMR spectroscopy.

    abstract::The heme environmental structures of lactoperoxidase (LP) have been studied by the use of hyperfine-shifted proton NMR and optical absorption spectra. The NMR spectra of the enzyme in native and cyanide forms in H2O indicated that the fifth ligand of the heme iron is the histidyl imidazole with an anionic character an...


    pub_type: 杂志文章


    authors: Shiro Y,Morishima I

    更新日期:1986-10-07 00:00:00

  • Photoconversion of 7-dehydrocholesterol to vitamin D3 in synthetic phospholipid bilayers.

    abstract::The incorporation of 7-dehydrocholesterol into synthetic phospholipid bilayers altered the distribution of products after photolysis. In liposomes, the relative amounts of 7-dehydrocholesterol and lumisterol were elevated, and tachysterol was reduced from the levels observed in hexane solution. Z to E isomerization of...


    pub_type: 杂志文章


    authors: Yamamoto JK,Borch RF

    更新日期:1985-06-18 00:00:00

  • Peptidyl boronates inhibit Salmonella enterica serovar Typhimurium Lon protease by a competitive ATP-dependent mechanism.

    abstract::Lon is a homo-oligomeric ATP-dependent serine protease that functions in the degradation of damaged and certain regulatory proteins. This enzyme has emerged as a novel target in the development of antibiotics because of its importance in conferring bacterial virulence. In this study, we explored the mechanism by which...


    pub_type: 杂志文章


    authors: Frase H,Lee I

    更新日期:2007-06-05 00:00:00

  • NMR structure determination of a novel conotoxin, [Pro 7,13] alpha A-conotoxin PIVA.

    abstract::A high-resolution solution conformation of a novel conotoxin, [Pro 7,13] alpha A-conotoxin PIVA, GCCGSYPNAACHPCSCKDROSYCGQ-NH2, has been determined by two-dimensional 1H NMR methods and distance geometry calculations. The total of 324 NOE-derived interproton distance restraints including 33 long-range NOE restraints a...


    pub_type: 杂志文章


    authors: Han KH,Hwang KJ,Kim SM,Kim SK,Gray WR,Olivera BM,Rivier J,Shon KJ

    更新日期:1997-02-18 00:00:00

  • Association of botulinum neurotoxin serotypes a and B with synaptic vesicle protein complexes.

    abstract::Botulinum neurotoxins (BoNTs) elicit flaccid paralysis through cleavage of SNARE proteins within peripheral neurons. There are seven serotypes of the BoNTs, termed A-G, which differ in the SNARE protein and/or site that is cleaved. BoNTs are single-chain toxins that comprise an N-terminal zinc metalloprotease domain t...


    pub_type: 杂志文章


    authors: Baldwin MR,Barbieri JT

    更新日期:2007-03-20 00:00:00

  • Transmembrane topography of the mitochondrial oxoglutarate carrier assessed by peptide-specific antibodies and enzymatic cleavage.

    abstract::The folding of the peptide chain of the bovine heart oxoglutarate carrier in the inner mitochondrial membrane and in the membrane of reconstituted proteoliposomes has been investigated by enzymatic and immunochemical approaches using proteinase K and polyclonal site-directed antibodies, respectively. Two peptides corr...


    pub_type: 杂志文章


    authors: Bisaccia F,Capobianco L,Brandolin G,Palmieri F

    更新日期:1994-03-29 00:00:00

  • Direct evidence for GTP and GDP-Pi intermediates in microtubule assembly.

    abstract::Identification of the kinetic intermediates in GTP hydrolysis on microtubules and characterization of their assembly properties is essential in understanding microtubule dynamics. By using an improved glass filter assay that selectively traps microtubules with a dead time of 2 s and monitoring taxol-induced rapid asse...


    pub_type: 杂志文章


    authors: Melki R,Carlier MF,Pantaloni D

    更新日期:1990-09-25 00:00:00

  • Thermodynamics of ligand binding to trp repressor.

    abstract::The thermodynamics of L-tryptophan and operator DNA binding to the tryptophan repressor of Escherichia coli were analyzed by titration microcalorimetry and van't Hoff analysis of footprinting titrations, respectively. At 25 degrees C in 10 mM sodium phosphate, pH 7.6, and 0.1 M NaCl, the binding of L-tryptophan to the...


    pub_type: 杂志文章


    authors: Jin L,Yang J,Carey J

    更新日期:1993-07-20 00:00:00

  • DNA binding specificity of the basic-helix-loop-helix protein MASH-1.

    abstract::Despite the high degree of sequence similarity in their basic-helix-loop-helix (BHLH) domains, MASH-1 and MyoD are involved in different biological processes. In order to define possible differences between the DNA binding specificities of these two proteins, we investigated the DNA binding properties of MASH-1 by cir...


    pub_type: 杂志文章


    authors: Meierhan D,el-Ariss C,Neuenschwander M,Sieber M,Stackhouse JF,Allemann RK

    更新日期:1995-09-05 00:00:00

  • Evidence for a slow tertiary relaxation in the reaction of tert-butyl isocyanide with horseradish peroxidase.

    abstract::The kinetics of tert-butyl isocyanide binding to the heme protein horseradish peroxidase (HRP) at 22 degrees C was examined on all time scales, from minutes to picoseconds, in aqueous borate buffer at pH 9.08. Unlike myoglobin (Mb) or hemoglobin, HRP shows two bimolecular ligand binding processes. For comparison, bind...


    pub_type: 杂志文章


    authors: Bandyopadhyay D,Walda KN,Grogan TM,Magde D,Traylor TG,Sharma VS

    更新日期:1996-02-06 00:00:00

  • Structure and function of PA4872 from Pseudomonas aeruginosa, a novel class of oxaloacetate decarboxylase from the PEP mutase/isocitrate lyase superfamily.

    abstract::Pseudomonas aeruginosa PA4872 was identified by sequence analysis as a structurally and functionally novel member of the PEP mutase/isocitrate lyase superfamily and therefore targeted for investigation. Substrate screens ruled out overlap with known catalytic functions of superfamily members. The crystal structure of ...


    pub_type: 杂志文章


    authors: Narayanan BC,Niu W,Han Y,Zou J,Mariano PS,Dunaway-Mariano D,Herzberg O

    更新日期:2008-01-08 00:00:00

  • Structural characteristics of heparan sulfates with varying sulfate contents.

    abstract::Structural properties of heparan sulfate preparations from hog mucosa and beef lung sources were obtained by application of Smith degradation and nitrous acid reactions. Products formed by these reactions indicated that most of the iduronic acid present in these mucopolysaccharides is ester sulfated, whereas N-sulfate...


    pub_type: 杂志文章


    authors: Cifonelli JA,King JA

    更新日期:1977-05-17 00:00:00

  • Reactivity of fibrinogen and fibrinopeptide A containing fibrinogen fragments with antisera to fibrinopeptide A.

    abstract::Two antisera used in the radioimmunoassay for human fibrinopeptide A (FPA) which appear to have different immunochemical specificities have been tested for cross-reactivity with fibrinogen and with three fragments of fibrinogen which contain the FPA sequence. These fragments were the three-chain, NH2-terminal disulfid...


    pub_type: 杂志文章


    authors: Canfield RE,Dean J,Nossel HL,Butler VP Jr,Wilner GD

    更新日期:1976-03-23 00:00:00

  • Quantification of quaternary structure stability in aggregation-prone proteins under physiological conditions: the transthyretin case.

    abstract::The quaternary structure stability of proteins is typically studied under conditions that accelerate their aggregation/unfolding processes on convenient laboratory time scales. Such conditions include high temperature or pressure, chaotrope-mediated unfolding, or low or high pH. These approaches have the limitation of...


    pub_type: 杂志文章


    authors: Robinson LZ,Reixach N

    更新日期:2014-10-21 00:00:00

  • Immunophilin AtFKBP13 sustains all peptidyl-prolyl isomerase activity in the thylakoid lumen from Arabidopsis thaliana deficient in AtCYP20-2.

    abstract::The physiological roles of immunophilins are unclear, but many possess peptidyl-prolyl isomerase (PPIase) activity, and they have been found in all organisms examined to date, implying that they are involved in fundamental, protein-folding processes. The chloroplast thylakoid lumen of the higher plant Arabidopsis thal...


    pub_type: 杂志文章


    authors: Edvardsson A,Shapiguzov A,Petersson UA,Schröder WP,Vener AV

    更新日期:2007-08-21 00:00:00

  • N-glycosyl bond formation catalyzed by human alkyladenine DNA glycosylase.

    abstract::The removal of damaged bases by DNA glycosylases is thought to be effectively irreversible, because of an overall equilibrium that favors hydrolysis over synthesis of the N-glycosyl bond. Surprisingly, human alkyladenine DNA glycosylase (AAG) can make damaged DNA by catalyzing formation of an N-glycosyl bond between 1...


    pub_type: 杂志文章


    authors: Admiraal SJ,O'Brien PJ

    更新日期:2010-10-26 00:00:00

  • Erythrocyte band 3 protein: evidence for multiple membrane-crossing segments in the 17 000-dalton chymotryptic fragment.

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    pub_type: 杂志文章


    authors: Jennings ML,Nicknish JS

    更新日期:1984-12-18 00:00:00

  • Access to the active site of periplasmic nitrate reductase: insights from site-directed mutagenesis and zinc inhibition studies.

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    pub_type: 杂志文章


    authors: Dementin S,Arnoux P,Frangioni B,Grosse S,Léger C,Burlat B,Guigliarelli B,Sabaty M,Pignol D

    更新日期:2007-08-28 00:00:00

  • The solution structure of clip domains from Manduca sexta prophenoloxidase activating proteinase-2.

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    pub_type: 杂志文章


    authors: Huang R,Lu Z,Dai H,Velde DV,Prakash O,Jiang H

    更新日期:2007-10-16 00:00:00

  • MAP4 is the in vivo substrate for CDC2 kinase in HeLa cells: identification of an M-phase specific and a cell cycle-independent phosphorylation site in MAP4.

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    pub_type: 杂志文章


    authors: Ookata K,Hisanaga S,Sugita M,Okuyama A,Murofushi H,Kitazawa H,Chari S,Bulinski JC,Kishimoto T

    更新日期:1997-12-16 00:00:00

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    pub_type: 杂志文章


    authors: Yang Y,Qin Y,Ding Q,Bakhtina M,Wang L,Tsai MD,Zhong D

    更新日期:2014-08-26 00:00:00

  • Redesign of the proton-pumping machinery of cytochrome c oxidase: proton pumping does not require Glu(I-286).

    abstract::One of the putative proton-transfer pathways leading from solution toward the binuclear center in many cytochrome c oxidases is the D-pathway, so-called because it starts with a highly conserved aspartate [D(I-132)] residue. Another highly conserved amino acid residue in this pathway, glutamate(I-286), has been indica...


    pub_type: 杂志文章


    authors: Aagaard A,Gilderson G,Mills DA,Ferguson-Miller S,Brzezinski P

    更新日期:2000-12-26 00:00:00