Recombinant molecule encoding human protease nexin

• US 5,278,049 A
• Filed: 03/13/1987
• Issued: 01/11/1994
• Est. Priority Date: 06/03/1986
• Status: Expired due to Fees

• Current Assignee: Incyte Pharmaceuticals, Inc., University of Kansas
• Original Assignee: Incyte Pharmaceuticals, Inc.
• Inventors: Baker, Joffre B., Scott, Randy W.
• Primary Examiner(s): Ellis, Joan
• Application Number:
  US07/025,450
• Time in Patent Office: 2,496 Days
• Field of Search: 435/68, 435/70, 435/172.3, 435/212, 435/219, 435/69.1, 435/71.1, 435/91, 435/252.33, 435/235.1, 435/320.1, 435/240.2, 435/255, 435/256, 536/27, 530/350
• US Class Current: 435/69.2
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/8121 Serpins

Abstract

DNA segments encoding two slightly different protease nexin I forms (PN-Iα and PN-Iβ) are cloned and expressed to provide practical quantities of PN-I for diagnostic and therapeutic use. PN-I is a serine protease inhibitor useful in controlling conditions mediated by proteolytic activity.

Claims

1. An isolated DNA molecule which encodes mature protease nexin-I (PN-I) alpha of FIG. 3 or PN-I beta of FIG. 4 or encodes a PN-I protein encoded by a gene which is a naturally occurring allelic variant of the gene encoding PN-I alpha or PN-I beta.

2. The DNA molecule of claim 1 which further includes DNA encoding a signal sequence which is operably linked to said mature PN-1α

or PN-1β

.

3. The DNA molecule of claim 2 wherein the linked DNA encodes the signal sequence of mature human PN-I.

4. A probe which is capable of hybridizing to mammalian cDNA that encodes PN-I and which is selected from the group consisting of

(1) the Sau3AI fragment of PN-18;

(2) the 750 bp EcoRI-Bg1II fragment of PN-18;

(3) the 14-mer of FIG. 1a; and

(4) the 36-mer of FIG. 1b.

5. An expression vector comprising a DNA encoding a protease nexin I (PN-I) having the amino acid sequence of PN-I alpha of FIG. 3 or PN-I beta of FIG. 4, or having an amino acid sequence encoded by a gene which is a naturally occurring allelic variant of that encoding said PN-I alpha or PN-I beta

wherein said expression vector is capable, when transformed into a host cell, of expressing said PN-I encoding DNA.

6. The expression vector of claim 5 wherein the PN-I encoding DNA includes DNA encoding an operably linked signal sequence.

7. A unicellular host cell transformed with the expression vector of claim 5.

8. The host cell of claim 7 wherein said host cell is a mammalian cell and wherein said vector comprises an SV40-derived promoter.

9. A method of producing PN-I alpha or PN-I beta which comprises culturing cells transformed with the expression system of claim 5 under conditions which induce the expression of said PN-I encoding DNA and

recovering the PN-I protein produced.