Phenylalanyl-tRNA Synthetase from Baker'Yeast: Structural Organization of the Enzyme and Its Complex with tRNAPhe as Determined by X-Ray Small-Angle Scattering

1979 ◽  
Vol 34 (1-2) ◽  
pp. 20-26 ◽  
Author(s):  
Ingrid Pilz ◽  
Karin Goral ◽  
Friedrich v. d. Haar
Keyword(s):  
Molecular Weight ◽  
Small Angle ◽  
Quaternary Structure ◽  
Trna Synthetase ◽  
Distribution Functions ◽  
Maximum Diameter ◽  
Radius Of Gyration ◽  
X Ray ◽  
Angle Scattering ◽  
The Difference

Abstract The quaternary structure of the phenylalanyl-tRNA synthetase and its complex with tRNAPhe was studied in dilute solutions by small angle X-ray scattering. For the free synthetase the radius of gyration was determined as 5.5 nm, the volume 523 nm3, the maximum diameter 17.5 nm and the molecular weight as 260 000 using an isopotential specific volume of 0.735. The overall shape could be best approximated by a flat cylinder with dimensions 18.2 nm X 11.5 nm X 4 nm ; the loose structure was approximated by building up the cylinder by spheres (diameter 4.2 nm).The corresponding parameters of the enzyme tRNA complex were the following: radius of gyration 5.9 nm, volume 543 nm 3, maximum diameter 21 nm and molecular weight 290 000. These parameters suggest an 1:1 complex, whereby it must be assumed that the tRNA molecule is attached in the extension of the longer axis. From the difference in the distance distribution functions of the free enzyme and the complex it is evident that we have to assume a change of conformation (contraction) of the enzyme upon the binding of the specific tRNA.

Investigation of Ribulose-1,5-bisphosphate Carboxylase-Oxygenase from Tobacco by Small Angle X-Ray Scattering: A Structural Model for the Enzyme in Solution

1988 ◽  
Vol 43 (5-6) ◽  
pp. 373-376 ◽  
Author(s):  
P. M. Abuja ◽  
I. Pilz
Keyword(s):  
Small Angle ◽  
Quaternary Structure ◽  
Higher Plants ◽  
Maximum Diameter ◽  
Radius Of Gyration ◽  
Bisphosphate Carboxylase ◽  
X Ray ◽  
X Ray Scattering ◽  
Inactive Form ◽  
Ray Scattering

The quaternary structure of ribulose-1,5-bisphosphate carboxylase/oxygenase from tobacco (Nicotiana tabacum) was investigated in solution by means of small angle X-ray scattering. The most important molecular parameters as the radius of gyration (Rg) and the maximum diameter (Dmax) were determined. Both the active and the inactive form of the enzyme were measured at 5 °C and at 20 °C. A more distinct difference in size could be detected between the inactive forms at these two temperatures (Rg = 4.80 nm (5 °C) and 4.68 nm (20 °C)) than between the active forms (Rg = 4.73 nm and 4.69 nm). The maximum diameters were determined to be 13.1 nm for the inactive form at 5 °C and 12.8 nm for the other forms. A model is proposed consisting of eight large and eight small subunits arranged in the way that seems to be typical for this enzyme in higher plants.

Information retrieval from X-ray small-angle scattering with polychromatic (`white') synchrotron radiation

1983 ◽  
Vol 16 (1) ◽  
pp. 42-46 ◽  
Author(s):  
O. Glatter ◽  
P. Laggner
Keyword(s):  
Synchrotron Radiation ◽  
Small Angle ◽  
Particle Shape ◽  
Structural Information ◽  
Small Angle Scattering ◽  
Radius Of Gyration ◽  
X Ray ◽  
Angle Scattering ◽  
Slit Length ◽  
Hinge Bending

The possibilities of obtaining structural information from X-ray small-angle scattering experiments with `white' polychromatic synchrotron radiation using line collimation are investigated by numerical simulation. Theoretical scattering curves of geometrical models were smeared with the appropriate wavelength distributions and slit-length functions, afflicted by statistical noise, and then evaluated by identical methods as normally used for experimental data, as described previously [program ITP; Glatter (1977). J. Appl. Cryst. 10, 415–421]. It is shown that even for a wavelength distribution of 50% half width, the information content is not limited to the parameters derived from the central part of the scattering curves, i.e. the radius of gyration and the zero-angle intensity, but also allows qualitative information on particle shape via the distance distribution function p(r). By a `hinge-bending model' consisting of two cylinders linked together at different angles it is demonstrated that changes in the radius of gyration amounting to less than 5% can be detected and quantified, and the qualitative changes in particle shape be reproduced.

Direct detection of the protein quaternary structure and denatured entity by small-angle scattering: guanidine hydrochloride denaturation of chaperonin protein GroEL

2002 ◽  
Vol 35 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Yuzuru Hiragi ◽  
Yasutaka Seki ◽  
Kaoru Ichimura ◽  
Kunitsugu Soda
Keyword(s):  
Small Angle ◽  
Quaternary Structure ◽  
Direct Detection ◽  
Guanidine Hydrochloride ◽  
Average Molecular Weight ◽  
Higher Order ◽  
Small Angle Scattering ◽  
Radius Of Gyration ◽  
Order Structure ◽  
Angle Scattering

A change in the higher-order structure of an oligomeric protein is directly detectable by small-angle scattering. A small-angle X-ray scattering (SAXS) study of the denaturation process of the chaperonin protein GroEL by guanidine hydrochloride (GdnHCl) showed that the disappearance of the quaternary structure can be monitored by using a Kratky plot of the scattered intensities, demonstrating the advantage of the SAXS method over other indirect methods, such as light scattering, circular dichroism (CD), fluorescence and sedimentation. The collapse of the quaternary structure was detected at a GdnHCl concentration of 0.8 Mfor a solution containing ADP (adenosine diphosphate)/Mg2+(2 mM)/K+. From pairwise plots of the change in forward scattering intensityJ(0)/C(weight-average molecular weight) and thez-average (root mean square) radius of gyration against the GdnHCl concentration, the stability and nature of the denatured protein can be determined. The SAXS results suggest that the GroEL tetradecamer directly dissociates to the unfolded coil without going through a globular monomer state. The denatured ensemble is not a single unfolded monomer coil particle, but some mixture of entangled aggregates and a monomer of the coil molecules. Small-angle scattering is a powerful method for the detection and study of changes in quaternary and higher-order structures of oligomeric proteins.

Small Angle X-Ray Study on the Structure of Active and Inactive Ribulose-1,5-bisphosphate Carboxylase-Oxygenase from Spinach. Evidence for a Configurational Change

1987 ◽  
Vol 42 (9-10) ◽  
pp. 1089-1091 ◽  
Author(s):  
Ingrid Pilz ◽  
Erika Schwarz ◽  
Gour P. Pal ◽  
Wolfram Saenger
Keyword(s):  
Small Angle ◽  
Quaternary Structure ◽  
Structural Models ◽  
Alcaligenes Eutrophus ◽  
Nitrogen Atmosphere ◽  
Radius Of Gyration ◽  
Bisphosphate Carboxylase ◽  
Configurational Change ◽  
X Ray ◽  
X Ray Scattering

Small angle X-ray scattering studies on ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) from spinach reveal a configurational change in its quaternary structure upon the transition of the molecule from the activated form occurring in the presence of CO2 and Mg2+ to the deactivated form obtained when CO2 and Mg2+ are removed by extensive dialysis under nitrogen. Present structural models are comparable to models which were postulated previously for the same enzyme but isolated from the hydrogen bacterium Alcaligenes eutrophus [O. Meisen- berger, I. Pilz, B. Bowien, G. P. Pal, and W. Saenger, J. Biol. Chem. 259, 4463-4465 (1984)]. The radius of gyration is R = 47.5 ±0.2 nm for the active spinach Rubisco. Upon deactivation, R changes to 49.2 ± 0.2 nm, suggesting a more elongated quaternary structure. The observed differ­ence in deactivation behaviour in ambient and in nitrogen atmosphere indicates a higher affinity of this spinach enzyme to CO2 with respect to the same enzyme from Alcaligenes eutrophus.

Glucose isomerase from Streptomyces rubiginosus – potential molecular weight standard for small-angle X-ray scattering

2005 ◽  
Vol 38 (3) ◽  
pp. 555-558 ◽  
Author(s):  
Maciej Kozak
Keyword(s):  
Molecular Weight ◽  
Synchrotron Radiation ◽  
Small Angle ◽  
Glucose Isomerase ◽  
Radius Of Gyration ◽  
X Ray ◽  
X Ray Scattering ◽  
Streptomyces Rubiginosus ◽  
Molecular Weight Standard ◽  
Ray Scattering

Stability of solutions of glucose isomerase from Streptomyces rubiginosus on long-term storage and on exposure to synchrotron radiation has been studied by the small-angle X-ray scattering (SAXS) method. The values of the radii of gyration and forward scattering do not change significantly on storage and on exposure to synchrotron radiation. The mean value of the radius of gyration characterizing glucose isomerase is R G = 3.27 ± 0.02 nm. For comparison, a SAXS study of monodispersive and aggregated bovine serum albumin (BSA) has been carried out. The results show that glucose isomerase could be a more stable molecular weight standard for SAXS than BSA.

A comparison of X-ray small-angle scattering results to crystal structure analysis and other physical techniques in the field of biological macromolecules

1978 ◽  
Vol 11 (1) ◽  
pp. 39-70 ◽  
Author(s):  
O. Kratky ◽  
I. Pilz
Keyword(s):  
Crystal Structure ◽  
Structure Analysis ◽  
Small Angle ◽  
Quaternary Structure ◽  
Heavy Atom ◽  
Crystal Structure Analysis ◽  
Small Angle Scattering ◽  
Biological Macromolecules ◽  
X Ray ◽  
Angle Scattering

In principle, there exist two ways to contribute to structure determination of macromolecules by X-ray diffraction: (a) by analysing diffraction data obtained from the crystalline state, and (b) by interpretation of X-ray small-angle scattering from particles in solution.The brilliant achievements of X-ray crystal-structure analysis of macromolecules, initiated by the works of Perutz on heamoglobin and Kendrew on myoglobin, are well known and it is evident that its detailed elution of secondary, tertiary and quaternary structure cannot be matched by any other means. However, a number of necessary prerequisites for a successful application, as, for example, the availability of well-defined crystals and heavy atom labelled derivatives thereof to surmount the problem of phase determination are not always given.

Neutron and X-ray small-angle scattering with Fe-based metallic glasses

1988 ◽  
Vol 97 ◽  
pp. 227-230 ◽  
Author(s):  
P. Lamparter ◽  
S. Steeb ◽  
D.M. Kroeger ◽  
S. Spooner
Keyword(s):  
Metallic Glasses ◽  
Small Angle ◽  
Small Angle Scattering ◽  
X Ray ◽  
Angle Scattering

Dimensions de micelles mixtes de p-alkylbenzènesulfonates par diffusion central des rayons X

1977 ◽  
Vol 10 (1) ◽  
pp. 37-44 ◽  
Author(s):  
C. Cabos ◽  
P. Delord ◽  
J. Rouviere
Keyword(s):  
Small Angle ◽  
Small Angle Scattering ◽  
Micellar Solutions ◽  
Probable Structure ◽  
X Ray ◽  
Absolute Scale ◽  
Angle Scattering ◽  
Scattering Measurements ◽  
Two Component ◽  
Most Probable Structure

The structure of micellar solutions is determined from X-ray small-angle scattering measurements on an absolute scale. The most probable structure is chosen by comparison with spherical cylindrical and lamellar models. This method is applied to two-component micelles and it is possible to follow the variation of micellar dimensions when the concentration of each component is varying.

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