EPR study of a copper center in a single crystal of cytosine monohydrate

Copper is found incorporated into the crystal structure of cytosine monohydrate grown from aqueous solution of commercially available cytosine. Upon ionizing irradiation, the crystals exhibited the electron paramagnetic resonance (EPR) spectra characteristic of Cu(II) complex. Planar coordination bonding to the cupric ion, having three nitrogen atoms and an oxygen as ligands, is interpreted to bridge two cytosine molecules, replacing the two cytosine-cytosine hydrogen bonds present in pure crystals. The EPR signals are much stronger for crystals grown from the solutions to which small amount of copper powder were added.     

Postirradiation long-range energy transfer in a single crystal of cytosine monohydrate: an EPR study.

Thiocytosine molecules incorporated in the cytosine monohydrate crystal lattice act as traps for both electrons and holes. The radiation-induced cytosine ion radicals, C(+) and C(-), release their charge upon heating. The excess electrons and holes migrate long distances in the crystal lattice. The migration of holes has been demonstrated by the postirradiation, thermally activated accumulation of thiocytosine cation radicals, T(+), and the migration of electrons by formation of the S-centered radicals of an anionic nature. It is estimated that the migration length of the holes is at least 30 interbase distances, and the migration length of the electrons is more than 100 interbase distances. The selective formation of the cationic and anionic trap radicals, depending on the trap concentration, is discussed in terms of differences between the migration of electrons and holes.     

Orientational order and dynamics of hydration water in a single crystal of bovine pancreatic trypsin inhibitor.

The orientational order and dynamics of the water molecules in form II crystals of bovine pancreatic trypsin inhibitor (BPTI) are studied by (2)H NMR in the temperature range 6-50 degrees C. From the orientation dependence of the single crystal quadrupole splitting and linewidth, the principal components of the motionally averaged quadrupole interaction tensor and the irreducible linewidth components for the orthorhombic crystal are determined. With the aid of water orientations derived from neutron and x-ray diffraction, it is shown that the NMR data can be accounted for by a small number of highly ordered crystal waters, some of which have residence times in the microsecond range. Most of these specific hydration sites must be located at intermolecular contacts. The surface hydration layer that is also present in dilute solution is likely to be only weakly ordered and would then not contribute significantly to the splitting and linewidth from the protein crystal. To probe water dynamics on shorter time scales, the (2)H longitudinal relaxation dispersion is measured for a polycrystalline BPTI sample. The observed dispersion is dominated by rapidly exchanging deuterons in protein side chains, undergoing restricted rotational motions on a time scale of 10 ns.     

Electron paramagnetic resonance of single crystal deoxycobaltohemoglobin

Single crystals of horse ^CoHb were obtained by reduction of ^CoHb⁺ crystals with dithionite. Epr measurements showed that the g? and ^Coà tensors are both axial and share the same principal axis systems. Of the four subunits, the “heme” normals of C? and d? subunits ãb?plane 29 ± 1° from b?; they have the same orientation as the hemes in methemoglobin. The normals of “hemes” à and B? are 47 above the ãb? plane as compared to 16° in methemoglobin.     

Electron paramagnetic resonance of Cu2+:Mb single crystal. Conformational changes.

Copper introduced into met-myoglobin crystals occupies various sites as indicated by electron paramagnetic resonance parameters. Cu2+ (A) is probably liganded to histidine A10, lysine A14, and asparagine GH4 (Banaszak et al., 1965) and shows superhyperfine interaction with a single (imidazole) nitrogen. Cu2+ (B) and Cu2+ (C) correspond to other anisotropic sites described in less detail. Cu2+ (A) exhibits a transition to an isotropic form with a transition temperature of 40.5 degrees C. This transition indicates a conformational change in myoglobin and could correspond to a motion of A helix away from the GH section. The transition temperature is 7 degrees C higher than the one previously reported (Atanasov, 1971) for myoglobin in solution.     

Development of a single detector ring micro crystal element scanner: QuickPET II

This article describes a single ring version of the micro crystal element scanner (MiCES) and investigation of its spatial resolution imaging characteristics for mouse positron emission tomography (PET) imaging. This single ring version of the MiCES system, referred to as QuickPET II, consists of 18 MiCE detector modules mounted as a single ring in a vertical gantry. The system has a 5.76-cm transverse field of view and a 1.98-cm axial field of view. In addition to the scanner and data acquisition system, we have developed an iterative reconstruction that includes a model of the system's detector response function. Evaluation images of line sources and mice have been acquired. Using filtered backprojection, the resolution for a reconstructed line source has been measured at 1.2 mm full width at half maximum. F-18-2-fluoro-2-deoxyglucose mouse PET images are provided. The result shows that QuickPET II has the imaging characteristics to support high-resolution, static mouse PET studies using 18-F labeled compounds.     

Structure of a second crystal form of Bence-Jones protein Loc: strikingly different domain associations in two crystal forms of a single protein

We have determined the structure of the immunoglobulin light-chain dimer Loc in a second crystal form that was grown from distilled water. The crystal structure was determined to 2.8-A resolution; the R factor is 0.22. The two variable domains are related by local 2-fold axes and form an antigen binding "pocket". The variable domain-variable domain interaction observed in this crystal form differs from the one exhibited by the protein when crystallized from ammonium sulfate in which the two variable domains formed a protrusion (Chang et al., 1985). The structure attained in the distilled water crystals is similar to, but not identical with, the one observed for the Mcg light-chain dimer in crystals grown from ammonium sulfate. Thus, two strikingly different structures were attained by this multisubunit protein in crystals grown under two different, commonly used, crystallization techniques. The quaternary interactions exhibited by the protein in the two crystal forms are sufficiently different to suggest fundamentally different interpretations of the structural basis for the function of this protein. This observation may have general implications regarding the use of single crystallographic determinations for detailed identification of structural and functional relationships. On the other hand, proteins whose structures can be altered by manipulation of crystallization conditions may provide useful systems for study of fundamental structural chemistry.     

High quality InP single crystal from Sumitomo electric

InP single crystals are primarily used as the conductive substrates for optical devices such as LEDs and lasers etc, in optoelectronics. Recently, active research for the improvement of MISFET, HEMT, HBT, RHET and OEICs has been demanding the development of a semi-insulating, SI, InP substrate on which these devices can be fabricated. The improvement of quality and reliability of these devices is being vigorously promoted, so high purity and low defect density are required of InP single crystal substrates. At Sumitomo Electric, the LEC method to grow InP single crystals with low defect density has been developed.     

Dynamic conformations of the CD38-mediated NAD cyclization captured in a single crystal

The extracellular domain of human CD38 is a multifunctional enzyme involved in the metabolism of two Ca²⁺ messengers: cyclic ADP-ribose and nicotinic acid adenine dinucleotide phosphate. When NAD is used as substrate, CD38 predominantly hydrolyzes it to ADP-ribose, with a trace amount of cyclic ADP-ribose produced through cyclization of the substrate. However, mutation of a key residue at the active site, E146, inhibits the hydrolysis activity of CD38 but greatly increases its cyclization activity. To understand the role of the residue E146 in the catalytic process, we determined the crystal structure of the E146A mutant protein with a substrate analogue, arabinosyl-2′-fluoro-deoxy-nicotinamide adenine dinucleotide. The structure captured the enzymatic reaction intermediates in six different conformations in a crystallographic asymmetric unit. The structural results indicate a folding-back process for the adenine ring of the substrate and provide the first multiple snapshots of the process. Our approach of utilizing multiple molecules in the crystallographic asymmetric unit should be generally applicable for capturing the dynamic nature of enzymatic catalysis.     

Structure and single crystal spectroscopy of Green Fluorescent Proteins

Usually, spectroscopic data on proteins in solution are interpreted at molecular level on the basis of the three-dimensional structures determined in the crystalline state. While it is widely recognized that the protein crystal structures are reliable models for the solution 3D structures, nevertheless it is also clear that sometimes the crystallization process can introduce some "artifacts" that can make difficult or even flaw the attempt to correlate the properties in solution with those in the crystalline state. In general, therefore, it would be desirable to identify some sort of control. In the case of the spectroscopic properties of proteins, the most straightforward check is to acquire data not only in solution but also on the crystals. In this regard, the Green Fluorescent Protein (GFP) is an interesting case in that a massive quantity of data correlating the spectroscopic properties in solution with the structural information in the crystalline state is available in literature. Despite that, a relatively limited amount of spectroscopic studies on single crystals of GFP or related FPs have been described. Here we review and discuss the main spectroscopic (in solution) and structural (in crystals) studies performed on the GFP and related fluorescent proteins, together with the spectroscopic analyses on various FPs members in the crystalline state. One main conclusion is that "in cristallo" spectroscopic studies are useful in providing new opportunities for gathering information not available in solution and are highly recommended to reliably correlate solution properties with structural features. This article is part of a Special Issue entitled: Protein Structure and Function in the Crystalline State.