Compositions and methods for protein structural determinations

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving viable micro-organism

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435 4, 435110, 435113, 435114, 4352536, 436 86, 436 15, C12Q 102, C12P 1314, C12P 1312, C12P 1310

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056270445

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BRIEF SUMMARY
FIELD OF THE INVENTION

This invention is concerned with the determination of the three-dimensional structure of biological macromolecules, especially proteins. In particular, it is concerned with novel compositions and methods for the determination, by NMR spectroscopy, of the three-dimensional structure of proteins expressed in cultures of mammalian or insect cells.


BACKGROUND OF THE INVENTION

For many years, there has been intense interest in the determination of the three-dimensional structure of biological macromolecules, particularly proteins. So-called "structure-function" studies have been carried out with a view to determining which structural features of a molecule, or class of molecules, are important for biological activity. Since the pioneering work of Nobel laureates, Perutz and coworkers on the structure of hemoglobin (Perutz, M. F. et al., Nature, 185, 416-422 (1960)) and Watson and Crick on the structure of DNA (Watson, J. D. and Crick, F. H. C.,Nature, 171, 737 (1953)), this field has been of major importance in the biological sciences.
More recently, there has evolved the concept of "rational drug design." This strategy for the design of drugs involves the determination of the three-dimensional structure of an "active part" of a particular biological molecule, such as a protein. The biological molecule may, for example, be a receptor, an enzyme, a hormone, or other biologically active molecule. Knowing the three-dimensional structure of the active site can enable scientists to design molecules that will block, mimic or enhance the natural biological activity of the molecule. (Appelt, K., et al., J. Med. Chem., 34, 1925 (1991)). The determination of the three-dimensional structure of biological molecules is therefore also of great practical and commercial significance.
The first technique developed to determine three-dimensional structures was X-ray crystallography. The structures of hemoglobin and DNA were both determined using this technique. X-ray crystallography involves bombarding a crystal of the material to be examined with a beam of X-rays which are refracted by the atoms of the ordered molecules in the crystal. The scattered X-rays are captured on a photographic plate, which is then developed using standard techniques. The diffracted X-rays are thus visualized as a series of spots on the plate, and from this pattern, the structure of the molecules in the crystal can be determined. For larger molecules, it is also necessary to crystallize the material with a heavy ion, such as ruthenium, in order to remove ambiguity due to phase differences.
More recently, another technique, nuclear magnetic resonance ("NMR") spectroscopy, has been developed to determine the three-dimensional structures of biological molecules, and particularly proteins. NMR spectroscopy was originally developed in the 1950's and has evolved into a powerful procedure for analyzing the structure of small compounds, such as those with a molecular weight of .ltoreq.1000 daltons. Briefly, the technique involves placing the material (usually in a suitable solvent) in a powerful magnetic field and irradiating it with a strong radio signal. The nuclei of the various atoms will align themselves with the magnetic field until energized by the radio signal. They then absorb this energy and re-radiate (resonate) it at a frequency dependent on i) the type of nucleus and ii) the chemical environment (determined largely by bonding) of the nucleus. Moreover, resonances can be transmitted from one nucleus to another, either through bonds or through three dimensional space, thus giving information about the environment of a particular nucleus and nuclei in the vicinity of it.
However, it is important to recognize that not all nuclei are NMR active. Indeed, not all isotopes of the same element are active. For example, whereas "ordinary" hydrogen, .sup.1 H, is NMR active, heavy hydrogen (deuterium), .sup.2 H, is not. Thus, any material that normally contains .sup.1 H hydrogen can be rendered "invisible" in the hydrogen NMR spectru

REFERENCES:
patent: 4182656 (1980-01-01), Ahnell et al.
patent: 5168225 (1992-12-01), Yamazaki et al.
patent: 5393669 (1995-02-01), Brown
G.J. Cox, J. Biol. Chem., 78 (1928), pp. 475-479.
J.D. Watson and F.H.C. Crick, Nature, 171 (1953), pp. 737-738.
M.F. Perutz et al., Nature, 185 (1960), pp. 416-422.
D.M. LeMaster and F.M. Richards, Analytical Biochemistry, 122 (1982), pp. 238-247.
L.E. Kay et al., Science, 249 (1990), pp. 411-414.
P.C. Driscoll et al., Nature, 353 (1991), pp. 762-765.
K. Appelt et al., Journ. of Med. Chem., 34 (1991), pp. 1925-1934.
V.L. Hsu and I.M. Armitage, Biochemistry, 31 (1992), pp. 12778-12784.
A.P. Hansen et al., Biochemistry, 31 (1992), p. 12713-12718.
S.J. Archer et al., Biochemistry, 32 (1993), pp. 1152-1163.
S.J. Archer et al., Biochemistry, 32 (1993), pp. 1164-1171.
Patents Abstracts of Japan, vol. 14, No. 503, issued Nov. 2, 1990, Nakamura et al., "Method of Analyzing Protein Structure", 20 Aug. 1990, Abs. grp. No. P1126, ABS Abs of JP 01-29261, see entire abstract.
Derwent Abstract of Japanese Patent JPO4046143, issued Feb. 17, 1992, Hitachi KK, "Synthesis of isotope multi-labelled aminoacid in improved yield-by reacting aldehyde(s) ammonium . . . hydrolyzing", see entire abstract.

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