Abstract:
:The secretory pro-hormone chromogranin A (CHGA) is densely packed into storage granules along with catecholamines, playing a catalytic role in granule biogenesis. 3-Dimensional structural data on CHGA are lacking. We found a superfamily structural homology for CHGA in the tropomyosin family of alpha-helical coiled-coils, even in mid-molecule regions where primary sequence identity is only modest. The assignment was confirmed by an independent algorithm, suggesting approximately 6-7 such domains spanning CHGA. We provide additional physiochemical evidence (chromatographic, spectral, microscopic) consistent with this unusual structure. Alpha-helical secondary structure (at up to approximately 45%) was confirmed by circular dichroism. CHGA molecular mass was estimated by MALDI-TOF mass spectrometry at approximately 50 kDa and by denaturing gel filtration at approximately 50-61 kDa, while its native Stokes radius was approximately 84.8 A, as compared to an expected approximately 30 A; the increase gave rise to an apparent native molecular weight of approximately 578 kDa, also consistent with the extended conformation of a coiled-coil. Small-angle X-ray scattering (SAXS) on CHGA in solution best fit an elongated cylindrical conformation in the monodisperse region with a radius of gyration of the rod cross-section (Rt) of approximately 52 A, compatible with a coiled-coil in the hydrated, aqueous state, or a multimeric coiled-coil. Electron microscopy with negative staining revealed an extended, filamentous CHGA structure with a diameter of approximately 94 +/- 4.5 A. Extended, coiled-coil conformation is likely to permit protein "packing" in the secretory granule at approximately 50% higher density than a globular/spherical conformation. Natural allelic variation in the catestatin region was predicted to disrupt the coiled-coil. Chromaffin granule ultrastructure revealed a approximately 108 +/- 6.3 A periodicity of electron density, suggesting nucleation of a binding complex by the CHGA core. Inhibition of CHGA expression, by siRNA, disrupted regulated secretory protein traffic by approximately 65%, while targeted ablation of the CHGA gene in the mouse reduced chromaffin granule cotransmitter concentrations by approximately 40-80%. These results suggest new roles for secretory protein tertiary structure in hormone and transmitter storage, with implications for secretory cargo condensation (or dense core "packing" structure) within the regulated pathway.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Mosley CA,Taupenot L,Biswas N,Taulane JP,Olson NH,Vaingankar SM,Wen G,Schork NJ,Ziegler MG,Mahata SK,O'Connor DTdoi
10.1021/bi700704rsubject
Has Abstractpub_date
2007-09-25 00:00:00pages
10999-1012issue
38eissn
0006-2960issn
1520-4995journal_volume
46pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::A five-residue loop structure in basic fibroblast growth factor (FGF-2) which extends from amino acid residue 118 to residue 122 was replaced, by cassette mutagenesis, with the corresponding seven-residue loop structure from the structural homologue acidic fibroblast growth factor (FGF-1) or the corresponding five-res...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00003a004
更新日期:1995-01-24 00:00:00
abstract::Copper-zinc superoxide dismutase (CuZnSOD) acquires its catalytic copper ion through interaction with another polypeptide termed the copper chaperone for SOD. Here, we combine X-ray crystallographic and analytical ultracentrifugation methods to characterize rigorously both truncated and full-length forms of apo-LYS7, ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992716g
更新日期:2000-04-04 00:00:00
abstract::P700, the primary electron donor of photosystem I is an asymmetric dimer made of one molecule of chlorophyll a' (P(A)) and one of chlorophyll a (P(B)). While the carbonyl groups of P(A) are involved in hydrogen-bonding interactions with several surrounding amino acid side chains and a water molecule, P(B) does not eng...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049515j
更新日期:2004-07-06 00:00:00
abstract::The NO adducts of leghemoglobin (Lb) are implicated in biological processes, but only the adduct with ferrous Lb (Lb(II)NO) has been characterized previously. We report the first characterization of ferric nitrosylleghemoglobin (Lb(III)NO) and XAS experiments performed on frozen aqueous solutions of Lb(II)NO and Lb(II...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990730n
更新日期:1999-12-14 00:00:00
abstract::7,8-diaminopelargonic acid (DAPA) synthase (EC 2.6.1.62) is a pyridoxal phosphate (PLP)-dependent transaminase that catalyzes the transfer of the alpha-amino group from S-adenosyl-L-methionine (SAM) to 7-keto-8-aminopelargonic acid (KAPA) to form DAPA in the antepenultimate step in the biosynthesis of biotin. The wild...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026339a
更新日期:2002-10-22 00:00:00
abstract::Resonance Raman vibrational spectra of the Pr, lumi-R, and Pfr forms of phytochrome have been obtained using low-temperature trapping and room temperature flow techniques in conjunction with shifted-excitation Raman difference spectroscopy (SERDS). The Pr to lumi-R photoconversion exhibits a thermal barrier and is com...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962175k
更新日期:1996-12-17 00:00:00
abstract::There is increasing evidence that a class of cell membrane glycolipids, gangliosides, can mediate the fibrillogenesis and toxicity of Alzheimer's disease amyloid-beta peptide (Abeta). Using lipid monolayers and vesicles as model membranes, we measured the insertion of Abeta into 1,2-dipalmitoyl-sn-glycero-3-phosphocho...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062177x
更新日期:2007-02-20 00:00:00
abstract::Resonance Raman and UV-visible absorption spectra show that nickel(II) microperoxidase-11 (NiMP-11) is four-coordinate in aqueous solution in the pH range from 1.0 to 13.0. In aqueous solutions of NiMP-11 in the absence of cetyltrimethylammonium bromide (CTAB), NiMP-11 is aggregated. In CTAB micellar solutions, where ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982332a
更新日期:1999-03-02 00:00:00
abstract::Carboxyl ester lipase (CEL; EC 3.1.1.13) hydrolyzes cholesteryl esters and retinyl esters in vitro. In vivo, pancreatic CEL is thought to liberate cholesterol and retinol from their esters prior to absorption in the intestine. CEL is also a major lipase in the breast milk of many mammals, including humans and mice, an...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981679a
更新日期:1999-03-30 00:00:00
abstract::Thermodynamic and structural evidence indicates that the DNA binding domains of lac repressor (lacI) exhibit significant conformational adaptability in operator binding, and that the marginally stable helix-turn-helix (HTH) recognition element is greatly stabilized by operator binding. Here we use circular dichroism a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0270992
更新日期:2003-02-25 00:00:00
abstract::Seven mutant LH1 polypeptides of Rhodobactor sphaeroides have been isolated, and their behaviors in in vitro reconstitution of LH1 and its subunit complex have been characterized. Two mutants were selected to address the increased stability of the subunit complex of Rb. sphaeroides compared with that of Rhodobacter ca...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962386p
更新日期:1997-03-25 00:00:00
abstract::We have studied the effect of the binding of ribosomal protein S1 and initiation factor IF3 on the accessibility of nucleotide residues 584-1506 in the small subunit of the Escherichia coli ribosome. Protein S1 strongly decreases RNase V1 attack at G1164, in hairpin 40 of the 3' major domain, and weakly decreases DMS ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00112a007
更新日期:1991-12-03 00:00:00
abstract::The catalytic activity and inhibitor binding energetics of enzymes are often pH-dependent properties. Aspartic proteases comprise an important class of enzyme targets for structure-based drug design. We have performed a complete thermodynamic study of pepstatin binding to plasmepsin II, an aspartic proteinase found in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi971550l
更新日期:1997-12-23 00:00:00
abstract::The nuclear tRNA 3' processing activity from wheat has been characterized and partially purified. Several characteristics of the wheat nuclear 3' processing enzyme now allow this activity to be distinguished from its mitochondrial counterpart. The nuclear enzyme is an endonuclease, which we termed nuclear RNase Z. The...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992253e
更新日期:2000-02-29 00:00:00
abstract::The sterol (free and esterified cholesterol) transport activity of the small-intestinal brush border membrane was solubilized with the short-chain detergent diheptanoylphosphatidylcholine and reconstituted to an artificial membrane system (proteoliposomes). The resulting proteoliposomes were identified as unilamellar ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970625i
更新日期:1997-09-02 00:00:00
abstract::Polyadenylated RNA was isolated from nuclei of cultured Drosophila cells, Schneider's line 2, and used as a template to synthesize a complementary DNA probe. Hybridization experiments were performed to study the relationship between nuclear and cytoplasmic RNA. About two-thirds of the nuclear polyadenylated RNA sequen...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00656a026
更新日期:1976-06-01 00:00:00
abstract::Binding mechanisms of ADPR-transferase to restricted double-stranded DNA fragments of SV40 and pBR322 DNA were determined by nuclease protection techniques. Top and bottom strands of double-stranded DNA were identified by specific labeling with 32P. Protection against specific exonucleases identified binding of ADPR-t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00439a050
更新日期:1989-06-27 00:00:00
abstract::Myelin basic protein (MBP) binds to negatively charged lipids on the cytosolic surface of oligodendrocyte membranes and is most likely responsible for adhesion of these surfaces in the multilayered myelin sheath. It can also polymerize actin, bundle F-actin filaments, and bind actin filaments to lipid bilayers through...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0519194
更新日期:2006-01-17 00:00:00
abstract::High signal-to-noise Fourier transform infrared (FTIR) spectra of the 5-hydroxytryptamine (serotonin) receptor (5-HT(3)R) and the nicotinic acetylcholine receptor (nAChR) were obtained by microscope FTIR spectroscopy using micrometer-sized, fully hydrated protein films. Because this novel procedure requires only nanog...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035113k
更新日期:2003-12-02 00:00:00
abstract::The mitochondrial tyrosyl tRNA synthetase from Neurospora crassa (CYT-18 protein) is a bifunctional group I intron splicing cofactor. CYT-18 is capable of splicing multiple group I introns from a wide variety of sources by stabilizing the catalytically active intron structures. CYT-18 and mt TyrRSs from related fungal...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi401614h
更新日期:2014-03-04 00:00:00
abstract::The ras oncogene products (H-Ras, K-Ras, and N-Ras) have been regarded as some of the most promising targets for anticancer drug discovery because their activating mutations are frequently found in human cancers. Nonetheless, molecular targeted therapy for them is currently unavailable. Here, we report the discovery o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00680
更新日期:2018-09-11 00:00:00
abstract::The interactions of the ligand binding domain (LBD) of androgen receptor (AR) and the AR T877A mutant, found in prostate cancer, with peptides from coactivator and corepressor proteins or random phage display peptides were investigated using in vitro time-resolved fluorescence resonance energy transfer (TR-FRET). Inte...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061321b
更新日期:2007-01-23 00:00:00
abstract::The interaction between skeletal myosin subfragment 1 (S1) and filamentous actin was examined at various intermediate states of the actomyosin ATPase cycle by chemical cross-linking experiments. Reaction of the actin-S1 complex with 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide and N-hydroxysuccinimide generated pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980139a
更新日期:1998-06-09 00:00:00
abstract::Triple-helical DNA shows increasing potential for applications in the control of gene expression (including therapeutics) and the development of sequence-specific DNA-cleaving agents. The major limitation in this technology has been the requirement of homopurine sequences for triplex formation. We describe a simple ap...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00117a002
更新日期:1992-01-21 00:00:00
abstract::The cation complexation equilibria between ionophore A23187 and several alkaline earth and first transition series divalent cations have been investigated. Absorption and fluorescence spectroscopy were used to monitor the reactions which were studied in solutions of 80% methanol/water, at 25 degrees C, and under condi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00390a019
更新日期:1987-08-11 00:00:00
abstract::Mammalian phospholipase D (PLD) has been implicated in the cellular signal transduction pathways leading to diverse physiological events and known to be regulated by many cellular factors. To identify the proteins that interact with PLD, we performed a protein overlay assay with fractions obtained from the sequential ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi015700a
更新日期:2002-03-12 00:00:00
abstract::A highly sensitive and stable scanning microcalorimeter is employed in a reinvestigation of the effect of cholesterol on multilamellar suspensions of dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC). Below 20 mol % cholesterol the DPPC mixtures give heat-capacity curves each of which can...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00605a034
更新日期:1978-06-13 00:00:00
abstract::The five-membered nitrogen plus heteroatom rings known as azolines or in their oxidized form as azoles are very common in natural products and drugs. The oxidation of thiazoline to thiazole in the cyanobactin class of natural products is one of the several important transformations that are known to alter the biologic...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00835
更新日期:2018-10-16 00:00:00
abstract::Marijuana is the most widely abused illegal drug, and its spectrum of effects suggests that several receptors are responsible for the activity. Two cannabinoid receptor subtypes, CB1 and CB2, have been identified, but the complex pharmacological properties of exogenous cannabinoids and endocannabinoids are not fully e...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200010k
更新日期:2011-06-28 00:00:00
abstract::The enzyme endonuclease V initiates repair of deaminated DNA bases by making an endonucleolytic incision on the 3' side one nucleotide from a base lesion. In this study, we have used site-directed mutagenesis to characterize the role of the highly conserved residues D43, E89, D110, and H214 in Thermotoga maritima endo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi060512b
更新日期:2006-08-29 00:00:00