The receptor binding and proteolysis of Spike SARS-CoV-2 release its S2 subunit to rearrange catalyze viral-cell fusion. This deploys the fusion peptide for insertion into cell membranes targeted. We show that this transforms from intrinsic disorder in solution a wedge-shaped structure inserted bilayered micelles, according chemical shifts, 15N NMR relaxation, NOEs. globular fold three helices contrasts open, extended forms region observed electron density compact prefusion states. In...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTPhenotypic and genetic characterization of cytochrome c2-deficient mutants Rhodobacter sphaeroidesTimothy J. Donohue, Alastair G. McEwan, Steven Van Doren, Antony R. Crofts, Samuel KaplanCite this: Biochemistry 1988, 27, 6, 1918–1925Publication Date (Print):March 22, 1988Publication History Published online1 May 2002Published inissue 22 March 1988https://pubs.acs.org/doi/10.1021/bi00406a018https://doi.org/10.1021/bi00406a018research-articleACS...

Abstract In plants, light-dependent activation of de novo fatty acid synthesis (FAS) is partially mediated by acetyl-CoA carboxylase (ACCase), the first committed step for this pathway. However, it not fully understood how plants control FAS regulation to meet cellular demand acyl chains. We report here identification a gene family encoding three small plastidial proteins envelope membrane that interact with α-carboxyltransferase (α-CT) subunit ACCase and participate in an original mechanism...

Methods are described for producing an active amino‐terminal domain of tissue inhibitor metalloproteinases‐1 (N‐TIMP‐1) from inactive protein expressed as inclusion bodies in E. coli . Yields exceed 20 mg per litre bacterial culture. Activity measurements, CD spectroscopy and NMR the 15 N‐labeled show that it is fully active, homogeneous conformation suitable high‐resolution structural analysis. The affinity N‐TIMP‐1 matrix metalloproteinases 1, 2 3 6–8‐fold less than recombinant full‐length...

Abstract Stromelysin, a representative matrix metalloproteinase and target of drug development efforts, plays prominent role in the pathological proteolysis associated with arthritis secondarily that cancer metastasis invasion. To provide structural template to aid therapeutic inhibitors, we have determined medium‐resolution structure 20‐kDa complex human stromelysin's catalytic domain hydrophobic peptidic inhibitor using multinuclear, multidimensional NMR spectroscopy. This this zinc...

Summary Gram‐negative phytopathogenic bacteria, such as Pseudomonas syringae , deliver multiple effector proteins into plant cells during infection. It is hypothesized that certain and mammalian need to traverse the type III secretion system unfolded are delivered host inactive enzymes. We have previously identified cyclophilin Arabidopsis eukaryotic activator of AvrRpt2, a P. cysteine protease. Cyclophilins general folding catalysts possess peptidyl‐prolyl cis/trans isomerase (PPIase)...

The topology of the cytochrome b subunit bc1 complex from Rhodobacter sphaeroides has been examined by generating gene fusions with alkaline phosphatase. Gene were generated at random locations within fbcB encoding subunit. These fusion products expressed in Escherichia coli and screened for phosphatase activity on chromogenic plates. 33 in-frame which showed further characterized. junctions all those had a high specific clustered three regions polypeptide, thus these tentatively assigned as...

Acetyl-CoA carboxylase (ACCase) catalyzes the first committed step in de novo synthesis of fatty acids. The multisubunit ACCase chloroplast is activated by a shift to pH 8 upon light adaptation and inhibited 7 dark adaptation. Here, titrations with purified biotin attachment domain-containing (BADC) carboxyl carrier protein (BCCP) subunits from Arabidopsis indicated that they can competently independently bind (BC) but differ responses changes representing those plastid stroma during or...

FHA domains are phosphoThr recognition modules found in diverse signaling proteins, including kinase-associated protein phosphatase (KAPP) from Arabidopsis thaliana. The kinase-interacting domain (KI-FHA) of KAPP targets it to function as a negative regulator some receptor-like kinase (RLK) pathways important plant development and environmental responses. To aid the identification potential binding sites for KI-FHA domain, we predicted (i) structure representative KAPP-binding RLK, CLAVATA1,...

The ubiquinol:cytochrome c2 oxidoreductase (bc1 complex) of Rhodobacter sphaeroides consists four subunits. One these subunits, cytochrome c1, is the site interaction with c2, a periplasmic protein. In addition, sequences fbcC gene and c1 subunit that it encodes suggest protein should be located on side cytoplasmic membrane anchored to by single membrane-spanning alpha-helix at carboxyl-terminal end polypeptide. Site-directed mutagenesis was used alter codon for Gln228 stop codon. This...

Hsc70's expected binding site on helix II of the J domain T antigens appears to be blocked in its structure bound tumor suppressor pRb. We used NMR map where mammalian Hsc70 binds murine polyomavirus (PyJ). The ATPase unexpectedly has biggest effects peak positions C-terminal end III PyJ. protects PyJ from an uncharged paramagnetic probe chelated Gd(III), clearly suggesting interface. Effects conserved HPD loop and are smaller. results supported by a novel assay ATP hydrolysis showing that...

Matrix metalloproteinase (MMP)-12 (or metalloelastase) efficiently hydrolyzed the gelatinase-selective α1(V)436-447 fluorescent triple helical peptide (THP) when substrate was submicromolar. The sequence of this THP derived from collagen V, a component I fibrils. hemopexin domains MMP-12 and -9 each increased kcat/Km toward by decreasing Km, just as domain MMP-1 enhances its peptidase activity. Non-fluorescent α1(V) subtly perturbed amide NMR chemical shifts not only in active site cleft but...

The pregnancy-associated glycoproteins (PAGs) represent a complex group of putative aspartic peptidases expressed exclusively in the placentas species Artiodactyla order. ruminant PAGs segregate into two classes: 'ancient' and 'modern' PAGs. Some modern possess alterations catalytic center that are predicted to preclude their ability act as peptidases. ancient contrast thought be peptidases, although no proteolytic activity has been described for these members. aim present study was...

Surfaces of the 173 residue catalytic domain human matrix metalloproteinase 3 (MMP-3(ΔC)) affected by binding N-terminal, 126 inhibitory TIMP-1 (N-TIMP-1) have been investigated using an amide-directed, NMR-based approach. The interface was mapped a novel method that compares amide proton line broadening paramagnetic Gd−EDTA in presence and absence partner. results are consistent with X-ray model complex MMP-3(ΔC) (Gomis-Rüth et al. (1997) Nature 389, 77−81). Residues Tyr155, Asn162, Val163,...

Evidence is presented that binding isotherms, simple or biphasic, can be extracted directly from noninterpreted, complex 2D NMR spectra using principal component analysis (PCA) to reveal the largest trend(s) across series. This approach renders peak picking unnecessary for tracking population changes. In 1:1 binding, first captures isotherm NMR-detected titrations in fast, slow, and even intermediate mixed exchange regimes, as illustrated phospholigand associations with proteins. Although...