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Journal Issue - Volume 9 Issue 4 (2000)

Abstract The structural study of peptides belonging to the terminal domains of histone H1 can be considered as a step toward the understanding of the function of H1 in chromatin. The conformational properties of the peptide Ac‐EPKRSVAFKKT KKEVKKVATPKK (CH‐1), which belongs to the C‐terminal domain of histone Hl° (residues 99–121) and is adjacent to the central globular domain of the protein, were examined by means of 1H‐NMR and...

Abstract The aim of the present study is to understand the structural features responsible for the lethal activity of snake venom cardiotoxins. Comparison of the lethal potency of the five cardiotoxin isoforms isolated from the venom of Taiwan cobra (Naja naja atra) reveals that the lethal potency of CTX I and CTX V are about twice of that exhibited by CTX II, CTX III, and CTX IV. In the present study, the solution structure of CTX V has been...

Abstract We have reported variants of the M13 bacteriophage major coat protein (P8) that enable high copy display of monomeric and oligomeric proteins, such as human growth hormone and steptavidin, on the surface of phage particles (Sidhu SS, Weiss GA, Wells JA. 2000. High copy display of large proteins on phage for functional selections. J Mol Biol 296:487–495). Here, we explore how an optimized P8 variant (opti‐P8) could evolve the ability to...

Abstract Physicochemical properties are potentially useful in predicting functional differences between aligned protein subfamilies. We present a method that considers physicochemical properties from ancestral sequences predicted to have given rise to the subfamilies of interest by gene duplication. Comparison between two map kinases subfamilies, p38 and ERK, revealed a region that had an excess of change in properties after gene...

Abstract The backbone dynamics of the four‐helical bundle cytokine leukemia inhibitory factor (LIF) have been investigated using 15N NMR relaxation and amide proton exchange measurements on a murine—human chimera, MH35‐LIF. For rapid backbone motions (on a time scale of 10 ps to 100 ns), as probed by 15N relaxation measurements, the dynamics parameters were calculated using the model‐free formalism incorporating the model selection approach....

  • Oxygen binding by α(Fe 2 hemoglobin crystals

  • Stefano Bruno, Stefano Bettati, Michele Manfredini, Andrea Mozzarelli, Martino Bolognesi, Daniela Deriu, Camillo Rosano, Antonio Tsuneshige, Takashi Yonetani, Eric R. Henry
  • Published in Wiley Interscience on Dec 31, 2008
  • DOI: 10.1110/ps.9.4.683 (p 683-692)

Abstract Oxygen binding by hemoglobin fixed in the T state either by crystallization or by encapsulation in silica gels is apparently noncooperative. However, cooperativity might be masked by different oxygen affinities of α and β subunits. Metal hybrid hemoglobins, where the noniron metal does not bind oxygen, provide the opportunity to determine the oxygen affinities of α and β hemes separately. Previous studies have characterized...

  • staphylococcus carnosus at high pressure

  • Hans Robert Kalbitzer, Adrian Görler, Hua Li, Peter V. Dubovskii, Wolfgang Hengstenberg, Claudia Kowolik, Hiroaki Yamada, Kazuyuki Akasaka
  • Published in Wiley Interscience on Dec 31, 2008
  • DOI: 10.1110/ps.9.4.693 (p 693-703)

Abstract The pressure‐induced changes in 15N enriched HPr from Staphylococcus carnosus were investigated by two‐dimensional (2D) heteronuclear NMR spectroscopy at pressures ranging from atmospheric pressure up to 200 MPa. The NMR experiments allowed the simultaneous observation of the backbone and side‐chain amide protons and nitrogens. Most of the resonances shift downfield with increasing pressure indicating generalized pressure‐induced...

Abstract Human infection with Toxoplasma gondii is an important cause of morbidity and mortality. Protozoan parasites such as T. gondii are incapable of de novo purine biosynthesis and must acquire purines from their host, so the purine salvage pathway offers a number of potential targets for antiparasitic chemotherapy. In T. gondii tachyzoites, adenosine is the predominantly salvaged purine nucleoside, and thus adenosine kinase is a key enzyme in the ...

Abstract The bacterial toxin colicin E9 is secreted by producing Escherichia coli cells with its 9.5 kDa inhibitor protein Im9 bound tightly to its 14.5 kDa C‐terminal DNase domain. Double‐ and triple‐resonance NMR spectra of the isolated DNase domain uniformly labeled with 13C/15N bound to unlabeled Im9 contain more signals than expected for a single DNase conformer, consistent with the bound DNase being present in more than one form. The...

Abstract The catalytic core of Escherichia coli DNA polymerase III contains three tightly associated subunits (α, €, and θ). The θ subunit is the smallest, but the least understood of the three. As a first step in a program aimed at understanding its function, the structure of the θ subunit has been determined by triple‐resonance multidimensional NMR spectroscopy. Although only a small protein, θ was difficult to assign fully...

Abstract We report an enigmatic peptide ligation reaction catalyzed by Glu‐specific Staphylococcus aureus V8 protease that occurs in neat aqueous solution around neutral pH utilizing a totally unprotected peptide substrate containing free α‐carboxyl and α‐amino groups. V8 protease catalyzed a chain of ligation steps between pH 6 and 8 at 4 °C, producing a gamut of covalent oligomers (dimer through octamer or higher) of a native protein segment...

Abstract At a time of the emergence of drug‐resistant bacterial strains, the development of antimicrobial compounds with novel mechanisms of action is of considerable interest. Perhaps the most promising among these is a family of antibacterial peptides originally isolated from insects. These were shown to act in a stereospecific manner on an as‐yet unidentified target bacterial protein. One of these peptides, drosocin, is inactive...

Abstract Surveys of protein crystal structures have revealed that amino acids show unique structural preferences for the N1, N2, and N3 positions in the first turn of the α‐helix. We have therefore extended helix‐coil theory to include statistical weights for these locations. The helix content of a peptide in this model is a function of N‐cap, C‐cap, N1, N2, N3, C1, and helix interior (N4 to C2) preferences. The partition function...

Abstract A simple method for growing protein crystals in the metastable zone using the vapor diffusion technique is described. The coverslips holding the hanging drops are transferred, after being incubated for some time at conditions normally giving many small crystals, over reservoirs at concentrations that normally yield clear drops. Fewer, much larger and better diffracting crystals are obtained, compared with conventional...

Abstract Using a cross‐linking approach, we recently demonstrated that radiolabeled peptides or misfolded proteins specifically interact in vitro with two luminal proteins in crude extracts from pancreas microsomes. The proteins were the folding catalysts protein disulfide isomerase (PDI) and PDIp, a glycosylated, PDI‐related protein, expressed exclusively in the pancreas. In this study, we explore the specificity of these proteins...

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