Bioluminescence of ()-Cypridina Luciferin with Luciferase.

Cypridina luciferin (CypL) is a marine natural product that functions as the luminous substrate for the enzyme luciferase (CypLase). CypL has two enantiomers, ()- and ()-CypL, due to its one chiral center at the -butyl moiety. Previous studies reported that ()-CypL or racemic CypL with CypLase produced light, but the luminescence of ()-CypL with CypLase has not been investigated.

Host-Dependent Producibility of Recombinant Luciferase With Glycosylation Defects.

luciferase (CLuc) is a secreted luminescent protein that reacts with its substrate (Cypridina luciferin) to emit light. CLuc is known to be a thermostable protein and has been used for various research applications, including imaging and high-throughput reporter assays. Previously, we produced a large amount of recombinant CLuc for crystallographic analysis.

Genetically engineered haemoglobin wrapped covalently with human serum albumins as an artificial O carrier.

We describe the synthesis and O affinity of genetically engineered human adult haemoglobin (rHbA) wrapped covalently with recombinant human serum albumins (rHSAs) as an artificial O carrier used for a completely synthetic red blood cell (RBC) substitute. Wild-type rHbA [rHbA(wt)] expressed in yeast species Pichia pastoris shows an identical amino acid sequence and three-dimensional structure to those of native HbA. It is particularly interesting that two orientations of the prosthetic haem group in rHbA(wt) were aligned by gentle heating in the natural form.

Quaternary Structure Analysis of a Hemoglobin Core in Hemoglobin-Albumin Cluster.

A core-shell ensemble of bovine hemoglobin (Hb) and human serum albumin (HSA) is an artificial O carrier as a red blood cell substitute. This protein particle is created by covalent wrapping of a carbonyl Hb with HSAs: Hb-HSA cluster, where Hb signifies the use of carbonyl Hb (relaxed (R) state conformation) as a starting material. The Hb-HSA cluster exhibits high O affinity and low cooperativity.

Core-shell protein clusters comprising haemoglobin and recombinant feline serum albumin as an artificial O carrier for cats.

This report describes the synthesis and structure of core-shell protein clusters comprising haemoglobin (Hb) at the centre and recombinant feline serum albumin (rFSA) at the exterior, named as haemoglobin-albumin clusters (Hb-rFSA). Specifically, we highlight their capability as an artificial O carrier that can be used as a red blood cell (RBC) substitute for cats, the most populous pet animal in the world. First, rFSA was expressed by genetic engineering using Pichia yeast.

Artificial Blood for Dogs.

There is no blood bank for pet animals. Consequently, veterinarians themselves must obtain "blood" for transfusion therapy. Among the blood components, serum albumin and red blood cells (RBCs) are particularly important to save lives.

Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1.

NAD+-reducing [NiFe] hydrogenases catalyze the oxidoreduction of dihydrogen concomitant with the interconversion of NAD+ and NADH. Here, the isolation, purification and crystallization of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1 are reported. Crystals of the NAD+-reducing [NiFe] hydrogenase were obtained within one week from a solution containing polyethylene glycol using the sitting-drop vapour-diffusion method and micro-seeding.

Resonance Raman spectral properties of FMN of bovine heart NADH:ubiquinone oxidoreductase suggesting a mechanism for the prevention of spontaneous production of reactive oxygen species.

A highly improved method for obtaining resonance Raman (RR) spectra provided spectra comparable to the best known flavoprotein spectra when the method was tested using bovine heart NADH:ubiquinone oxidoreductase (Complex I), a protein with a molecular mass of 1000 kDa, which causes the level of RR noise to be 1 order of magnitude higher than for most flavoproteins. The FMN RR band shift (1631/1633 cm(-1)) and the increase in the magnitude of the band at 1252 cm(-1) upon binding to Complex I suggest hydrogen bond formation involving one of the C=O groups [C(2)=O] of isoalloxazine to stabilize its quinoid form. This lowers the redox potential of FMN and the electron density of the O(2) binding site [a carbon atom, C(4a)] in the reduced form.

Crystal structure analysis of the translation factor RF3 (release factor 3).

The bacterial translational GTPases release factor RF3 promotes translation termination by recycling RF1 or RF2. Here, we present the crystal structures of RF3 complexed with GDP and guanosine 3',5'-(bis)diphosphate (ppGpp) at resolutions of 1.8 and 3.

Crystallization and preliminary X-ray analysis of a class II release factor RF3 from a sulfate-reducing bacterium.

Class II release factor 3 (RF3) from the sulfate-reducing bacterium Desulfovibrio vulgaris Miyazaki F, which promotes rapid dissociation of a class I release factor, has been overexpressed, purified and crystallized in complex with GDP at 293 K using the sitting-drop vapour-diffusion method. A data set was collected to 1.8 A resolution from a single crystal at 100 K using synchrotron radiation.