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G Zaccai

Publications and source records attributed to G Zaccai.

87 records · Page 5Linked to original sources
PubMed1980

Neutron scattering studies of lac repressor.

The lac repressor, a tetrameric protein of identical subunits [molecular weight (MW) 4 x 38,500], interacts specifically with the lac operator, preventing the expression of the structural genes of the lac operon. The presence of an inducer (such as isopropyl-beta-D-thiogalactoside; IPTG) which binds to the repressor, prevents operator binding by lowering the association constant by a factor of 10(3) (ref. 3). Genetic and biochemical analysis have shown that the major part--if not all--of the binding site for the lac operator is located in the 60 N-terminal residues of the protein. In certain conditions, limited trypsinolysis of the protein yields four N-terminal 'headpieces' each containing 51 or 59 residues, and a tetrameric core with full inducer binding activity. It was shown recently that this headpiece is able to bind nucleic acids, and interacts with the lac operator, giving the same pattern of sensitivity with respect to the methylation of the bases as does the intact repressor. We are studying the interaction of lac repressor with DNA by neutron scattering using contrast variation and discuss here measurements on the protein, its tryptic core and their complexes with IPTG. Our results demonstrate that the headpieces are located far (67 +/- 10 A) from the centre of mass of a somewhat elongated core, and that the inducer does not significantly change the radius of gyration of the protein.

Allosteric Regulation↗
PubMed1980

Bacteriorhodopsin is an inside-out protein.

Neutron scattering is particulary useful when parts of a structure can be deuterated. From Halobacterium halobium we have obtained, by biosynthetic incorporation, purple membranes in which all of the valines or all of the phenylalanines are present in deuterated form. Difference Fourier techniques permit a general assessment of the distribution of valine and phenylalanine in projections of the purple membrane structure. These show that valine is distributed toward the periphery of a single bacteriorhodopsin molecule, whereas phenylalanine is distributed toward its center. We use the facts that the amino acid sequence is known and that much of it can be assigned to the alpha helices of the bacteriorhodopsin structure to interpret our results. Comparison of our maps with the distribution of valine and phenylalanine around alpha-helical perimetrs establishes the distribution of other amino acids and leads to the conclusion that the charged and polar groups of the bacteriorhodopsin molecule tend to lie at the molecular interior, away from contact with lipid, while the nonpolar surfaces are directed outward, making contact with the lipid regions. Thus, the protein is "inside-out" compared with the organization of soluble proteins.

Bacteriorhodopsins↗
PubMed1977

A neutron investigation of yeast valyl-tRNA synthetase interaction with tRNAs.

A new way of studying RNA-protein complexes, using neutron small angle scattering in solution, is described and was applied in the case of the system, yeast valyl-tRNA synthetase, interacting with its cognate and non cognate yeast tRNAs. It was shown that, when limited amounts of tRNA (either cognate or non cognate) are added to valyl-tRNA synthetase, a complex consisting of two enzyme molecules and one tRNA molecule is first formed. It is subsequently dissociated to a one to one complex when more tRNA is present in the solution. The association curve shows a maximum for a molecular ratio, enzyme over tRNA, equal to 2.

Amino Acyl-tRNA Synthetases↗
PubMed1976

Direct methods for the analysis of lamellar neutron diffraction from oriented multilayers: a difference Patterson deconvolution approach.

We have described here a direct method for the solution of the phase problem for lamellar neutron diffraction from hydrated oriented multilayers of model membranes. As a result, the neutron-scattering profiles of such model membranes can be uniquely determined. We note that such methods can easily be extended to cases in which the thickness of the water layer w is somewhat greater than d/2 as long as it is substantially less than d [e.g., w less than 3d/4 would suffice, since deltaP(-d/4 less than or equal to x less than or equal to d/4) would contain only contributions from see article without overlapping contributions from see articles and to cases in which isotopic labeling is used instead of partial H2O-D2O exchange.

Cholesterol↗
PubMed1976

Some focusing techniques and their application to low-angle neutron scattering.

Both imaging and simple converging collimator systems can provide a focused neutron beam. Each system has been tested and appears suitable for exploitation. Imaging systems employing bent multilayer monochromators can be made simply by elastically bending a flat multilayer. The periodic spacing of the multilayer can be made to match that of the sample. The converging Soller slit provides a purely geometric means of prouducing a focused beam. Wavelength spread and distribution across the beam can be controlled separately. Soller-slit technology is further advanced than that of multilayers. The experiments sketched above demonstrate that converging or focused neutron beams suitable for low-angle scattering work can be produced with existing technology. Focusing incurs little or no loss of intensity and does not seriously distort the resolution function of a low-angel instrument. It permits a small spectrometer to make use of both advanced detector technology and large samples and thus to approach the capability of a much larger machine.

Neutrons↗
PubMed1975

Neutron diffraction studies on the location of water in lecithin bilayer model membranes.

Lamellar neutron diffraction from oriented multilayers of hydrated dipalmitoyl lecithin was phased by isomorphous H(2)O-D(2)O exchange and swelling techniques. Bound water sites were located in the polar head group region of the bilayer profile. A 6-A resolution structure based on the neutron scattering density profile is proposed for the bilayer. It is consistent with the electron density profile from x-ray diffraction.

Journal Article↗
PubMedDate not supplied

[Isolation and physical study of the 13S fragment of 16S RNA and its complex with ribosomal protein S4].

A fragment of E. coli 16S RNA has been obtained by its hydrolysis with pancreatic RNAase A coupled to Sepharose 4B. This fragment has a molecular weight of 170 000 and a sedimentation coefficient of 13S. It does not aggregate in solution and binds with the ribosomal protein S4. The 13S fragment and it complex with the protein S4 have been studied by different physical methods in the first place, by neutron scattering. It has been shown that this fragment is compact in solution. The radii of gyration of the fragment (50 +/- 3 A) and of the protein S4 within the complex (17 +/- 3 A) coincide, within limits of experimental error, with the radii of gyration for the free RNA fragment (47 +/- 2 A) and the free ribosomal protein S4 in solution (18 +/- 2 A). Hence, the conclusion is made that the compactness of the 13S fragment of the 16S RNA and the ribosomal protein S4 does not change at the complex formation. The compact 13S fragment of the 16S RNA is shown to be contrast matched in the H2O/D2O mixture containing 70% D2O which corresponds to its partial specific volume v equal to 0.537 cm3/g.

Chemical Phenomena↗