HOMEOSTATIC MULTIDOMAIN PROTEIN, AND USES FOR IT

• US 20120122107A1
• Filed: 11/16/2010
• Published: 05/17/2012
• Est. Priority Date: 11/16/2010
• Status: Abandoned Application

• Current Assignee: Jens Christian Jensenius, Soren Degn, Steffen Thiel
• Original Assignee: Aarhus Universitet (Ministeriet For Forskning Innovation OG Videregående UD- Dannelser)
• Inventors: Thiel, Steffen, Jensenius, Jens Christian, Degn, Søren
• Application Number:
  US12/947,751
• Publication Number:
  US 20120122107A1
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: G01N 2333/4703 Regulators; Modulating activity
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors

Abstract

The invention relates to the discovery and characterization of mannan binding lectin-associated protein (map44), a new protein that acts in the lectin pathway of complement activation.

Claims

1. An isolated polypeptide comprising:

(a) the amino acid sequence of SEQ ID NO 1;

(b) an amino acid sequence with at least about 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ. ID. NO. 1, wherein said polypeptide is capable of binding to mannan-binding lectin (MBL), H-ficolin, L-ficolin, or M-ficolin;

or

(c) a fragment of SEQ. ID. NO. 1 that is at least 50, 60, 70, 80, 90, or 100 amino acids in length, wherein said fragment is capable of competitively inhibiting MBL-associated serine proteases (MASPs) or the formation of the MBLIMASP or ficolin/MASP complex.

2. The isolated polypeptide of claim 1, further comprising a label or toxin joined thereto.

3. The isolated polypeptide of claim 1, wherein said polypeptide comprises SEQ ID NO 1.

4. The isolated polypeptide of claim 1, wherein said polypeptide comprises an amino acid sequence with at least about 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ. ID. NO. 1 and, wherein said polypeptide is capable of binding to mannan-binding lectin (MBL), H-ficolin, L-ficolin, or M-ficolin.

5. The isolated polypeptide of claim 1, wherein said polypeptide comprises a fragment of SEQ. ID. NO. 1 that is at least 50, 60, 70, 80, 90, or 100 amino acids in length and, wherein said fragment is capable of competitively inhibiting MBL-associated serine proteases (MASPs) or the formation of the MBLIMASP or ficolin/MASP complex.

6. The isolated polypeptide of claim 3, further comprising a label or toxin joined thereto.

7. An isolated nucleic acid that comprises:

(a) a nucleotide sequence that encodes SEQ ID NO 1;

(b) the nucleotide sequence of SEQ ID NO 2;

(c) a fragment of a nucleotide sequence of SEQ ID NO 2 that is at least about 50, 60, 70, 80, 90, 100, 125, 150, or 200 nucleotides in length;

or

(d) a nucleotide sequence with at least about 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ. ID. NO. 2, wherein the polypeptide encoded by said nucleic acid is capable of binding to mannan-binding lectin (MBL), H-ficolin, L-ficolin, or M-ficolin.

8. The isolated nucleic acid of claim 7, wherein said nucleic acid comprises a nucleotide sequence that encodes SEQ ID NO 1.

9. The isolated nucleic acid of claim 7, wherein said nucleic acid comprises the nucleotide sequence of SEQ ID NO 2.

10. The isolated nucleic acid of claim 7, wherein said nucleic acid comprises a fragment of a nucleotide sequence of SEQ ID NO 2 that is at least about 50, 60, 70, 80, 90, 100, 125, 150, or 200 nucleotides in length.

11. The isolated nucleic acid of claim 7, wherein said nucleic acid comprises a nucleotide sequence with at least about 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ. ID. NO. 2 and, wherein the polypeptide encoded by said nucleic acid is capable of binding to mannan-binding lectin (MBL), H-ficolin, L-ficolin, or M-ficolin.

12. The isolated nucleic acid of claim 7, further comprising a vector sequence.

13. The isolated nucleic acid of claim 8, further comprising a vector sequence.

14. The isolated nucleic acid of claim 9, further comprising a vector sequence.

15. The isolated nucleic acid of claim 12, wherein said vector is an expression vector.

16. The isolated nucleic acid of claim 7, further comprising a regulatory element linked operably thereto.

17. The isolated nucleic acid of claim 8, further comprising a regulatory element linked operably thereto.

18. The isolated nucleic acid of claim 9, further comprising a regulatory element linked operably thereto.

19. A method for identifying the presence of MAP44 in a biological sample, comprising

detecting the amount of a polypeptide comprising SEQ ID NO 1 or the amount of an RNA encoding SEQ ID NO 1 in a biological sample.

20. The method of claim 19, further comprising comparing the amount of the polypeptide comprising SEQ ID NO 1 or the amount of the RNA encoding SEQ ID NO 1 detected with a reference amount of MAP44 or an RNA encoding MAP44 or the amount of the polypeptide comprising SEQ ID NO 1 or the amount of the RNA encoding SEQ ID NO 1 detected in a second biological sample.