COMPOSITIONS, SYSTEMS AND METHODS FOR THE DIAGNOSIS, PREVENTION AND TREATMENT OF DISORDERS ASSOCIATED WITH AZETIDINE-2-CARBOXYLIC ACID

Abstract

Methods and systems for detecting azetidine-2-carboxylic acid (Aze) in food consumable by humans and animals are provided. Also provided are methods and systems for inactivating Aze in food and byproducts, as well as other methods for the diagnosis, prevention, and treatment of disorders associated with Aze.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

The disclosure can be better understood with reference to the following drawings. The components in the drawings are not necessarily to scale, emphasis instead being placed upon clearly illustrating the principles of the present disclosure. The drawings are described in greater detail in the description and examples below.

FIG. 1 illustrates the chemical structure of proline (left) and azetidine-2-carboxylic acid (Aze) (right).

FIG. 2 illustrates chromatographic peaks of (A) pure Aze, (B) liquid milk, and (C) liquid milk spiked with Aze, using a Hitachi L-8800 amino-acid analyzer.

FIG. 3 illustrates chromatographic peaks of two different varieties of yeast (Saccharomyces cerevisiae), samples A and B, showing the peaks of Aze in each. The analyses were done using a Beckman (Li) amino-acid analyzer.

FIG. 4 illustrates NMR spectra of 6 peptide derivatives of myelin basic peptide. Peptide 1 represents amino acids 95 to 103 of a wild-type human myelin basic protein (SEQ. ID. NO. 1). Peptides 2-6 (SEQ. ID. NOS. 2-6) are derivatives of peptide 1 with one or more proline residues replaced with Aze.

FIG. 5 illustrates a TOCSY two-dimensional proton spectra of peptide 1 (SEQ. ID. NO. 1).

FIG. 6 illustrates a ROESY two-dimensional proton spectra of peptide 1 (SEQ. ID. NO. 1).

Claims

1. A method comprising: detecting Aze in food consumable by humans or animals.

2. The method of claim 1, wherein the food is selected from at least one of: garden beets and sugar beets.

3. The method of claim 1, wherein the food is an animal product obtained from an animal fed materials containing Aze.

4. The method of claim 3, wherein the food is milk.

5. The method of claim 1, wherein the food is yeast or a food containing yeast.

6. The method of claim 3, wherein the food is selected from beef, pork, poultry, and eggs.

7. The method of claim 1, wherein detecting comprises detecting Aze using chromatography.

8. A method comprising: inactivating Aze in food consumable by humans or animals by heating the food containing Aze or an ingredient of food containing Aze to a temperature of about 100 degrees to 200 degrees, wherein the heat is effective to rupture the ring-structure of Aze.

9. The method of claim 8, comprising heating the food to a temperature of about 115 degrees to 180 degrees.

10. The method of claim 8, wherein the food comprises beet byproducts.

11. A method comprising: inactivating Aze in food consumable by humans or animals by contacting the food with an effective amount of Aze acetyltransferase, wherein the Aze acetyltransferase inactivates Aze by acetylation.

12. The method of claim 11, wherein the Aze acetyltransferase is selected from at least one of the following: acetyltransferase MPR1 from Saccharomyces cervisiae and acetyltransferase MPR2 from Saccharomyces cervisiae.

13. A method comprising: administering to a host who has recently consumed material containing Aze a therapeutically effective amount of a composition comprising proline, wherein the proline is effective to reduce the amount of Aze incorporated into host proteins.

14. The method of claim 13, wherein the proline comprises substantially pure L-proline.

15. The method of claim 13, wherein the composition further comprises a pharmaceutically acceptable carrier.

16. The method of claim 13, wherein the host proteins are selected from one or more of the following: myelin basic protein, collagen, hypoxia-inducible factor, profilins, ion channel proteins, vesicular glutamate transporters, and hemoglobins.

17. The method of claim 13, wherein the host is a pregnant female and wherein the proline is effective to reduce or prevent incorporation of Aze into fetal proteins.

18. The method of claim 17, wherein the proline is effective to reduce the risk of birth defects mediated by Aze misincorporation into fetal proteins.

19. A method of treating a disorder associated with Aze comprising: administering to a host in need of treatment for a disorder associated with Aze, a therapeutically effective amount of a composition comprising proline, wherein the proline is effective to reduce the amount of Aze incorporated into host proteins or to prevent additional incorporation of Aze into host proteins.

20. The method of claim 19, wherein the disorder is selected from at least one of the following: multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, systemic scleroses, mixed connective tissue disorders, and inflammatory myopathies and vasculitides.

21. The method of claim 19, wherein the disorder is multiple sclerosis.

22. The method of claim 19, wherein the disorder is an auto-immune disorder.

23. The method of claim 19, wherein the host proteins are selected from one or more of the following: myelin basic protein, collagen, hypoxia-inducible factor, profilins, ion channel proteins, vesicular glutamate transporters, and hemoglobins.

24. A method of detecting Aze in host polypeptides comprising: obtaining a sample from a host, wherein the sample comprises host polypeptides; andcontacting the host sample with a composition comprising at least one anti-Aze antibody, wherein the at least one anti-Aze antibody recognizes at least one host polypeptide having Aze in place of proline in the polypeptide sequence.

25. A method of detecting antibodies to Aze in a host comprising: providing at least one peptide comprising Aze, wherein the peptide is a derivative of a wild-type peptide having at least one proline residue replaced by Aze;providing a sample from a host, wherein the sample comprises host antibodies;contacting the host sample with the at least one peptide; anddetecting binding between the peptide and an antibody from the host sample, wherein binding indicates the presence of an antibody to the Aze-containing peptide.

26. The method of claim 25, wherein the host has multiple sclerosis and the peptide comprising Aze comprises a portion of the sequence of a myelin basic protein having a proline residue replaced by Aze.

27. A method of detecting antibodies to Aze comprising: providing a library of peptides comprising Aze in place of at least one proline residue;contacting the library of peptides with a composition comprising antibodies; anddetecting binding between the peptides and an antibody from the antibody composition, wherein binding indicates the presence of an antibody to the Aze-containing peptide.

28. The method of claim 27, wherein the library of peptides comprises at least one of the peptides of SEQ. ID. NOS. 2-8.

29. The method of claim 27, wherein the library of peptides comprises portions of one or more of the following human proteins: myelin basic protein, collagen, hypoxia-inducible factor, profilins, ion channel proteins, vesicular glutamate transporters, and hemoglobins, wherein the wild-type sequence of each protein portion comprises at least one proline residue, and wherein at least one proline of each peptide in the library has been replaced by Aze.

30. A polypeptide comprising a sequence selected from: SEQ. ID. NOS. 2-8.

31. A polypeptide consisting of a sequence selected from: SEQ. ID. NOS. 2-8.

32. An antibody capable of binding a polypeptide comprising a sequence selected from: SEQ. ID. NOS. 2-8.