Manganese superoxide dismutase cloning and expression in...

Drug – bio-affecting and body treating compositions – Enzyme or coenzyme containing – Oxidoreductases

Reexamination Certificate

Rate now

  [ 0.00 ] – not rated yet Voters 0   Comments 0

Details

C435S189000

Reexamination Certificate

active

06326003

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The production of the enzyme manganese human superoxide dismutase (hSODm) by recombinant DNA techniques is described. Both natural and modified enzymes are produced utilizing novel DNA constructs, plasmids and transformed microbial expression systems.
Superoxide dismutase (“SOD”) is in fact a variety of different enzymes found in most living organisms. One function in mammals is to destroy superoxide. Superoxide is a material naturally produced during phagocytosis and aerobic metabolism. The superoxide dismutases are characterized in families based on the metal ion associated with the enzyme, where the ions can be iron, manganese, copper, and copper and zinc. Superoxide dismutase, e.g., from bovine liver, has found clinical use, particularly as an anti-inflammatory agent in mammals including humans and to decrease tissue injury due to reperfusion (post-ischemic). Other utilities include scavenging superoxide anions due to exposure of a host to various superoxide-inducing agents, e.g. radiation, paraquat, etc.; prophylaxis or therapy for certain degenerative diseases, e.g., emphysema; food preservation; and the like.
It is therefore important that stable supplies of physiologically acceptable superoxide dismutase be made available, particularly for use in vivo as an anti-inflammatory agent or for other therapeutic purposes. For human application it would be preferable to employ the homologous enzyme to prevent or minimize possible immune response. By employing recombinant DNA techniques, there is the opportunity to produce products efficiently, which have the desired biological activities of superoxide dismutase, such as immunological and enzymatic activities.
2. Description of Relevant Publications
The primary structure of human liver manganese superoxide dismutase was described by Barra et al., J.B.C., 259:12595-12601, (1984). Either bacterial iron superoxide dismutase (FeSOD) or bacterial manganese superoxide dismutase (MnSOD) were shown to be required as a defense against oxygen toxicity by Carlioz et al., EMBO Journal, 5:623-630, (1986). The amino-terminal processing of methionine by yeast was shown by Tsunasawa et al., J.B.C., 260:5382-5391, (1985). Human copper/zinc superoxide dismutase was described by Hallewell, et al., Nucleic Acids Research, 13:2017-2034, (1985). A superoxide dismutase produced in
Serratia marcescens
is described in EPO application 0172577. An immobilized superoxide dismutase was described in EPO application 070656.
The amino acid sequence of human erythrocyte Cu—Zn superoxide dismutase was described in Jabusch et al.,
Biochemistry
(1980) 19:2310-2316 and Barra et al.,
FEBS Letters
(1980) 120:53-55. Bovine erythrocyte Cu—Zn SOD was described by Steinman et al.,
J. Biol. Chem
. (1974) 249:7326-7338. A SOD-1 (Cu—Zn SOD) cDNA clone is described by Lieman-Hurwitz et al.,
Proc. Natl. Acad. Sci. USA
(1982) 79:2808-2811.
SUMMARY OF THE INVENTION
Novel compositions are provided comprising nucleic acid sequences for the expression of polypeptides exhibiting the biological properties of human manganese superoxide dismutase (“hSODm”). Also provided are methods for producing such polypeptides employing recombinant DNA techniques and microorganism hosts. The subject polypeptides find use in vivo and in vitro in destroying superoxide.
Also provided is a novel modified hSODm. A modified DNA coding for hSODm with the first two amino acids, lysine and histidine, removed, resulted in a polypeptide with the third amino acid, serine, positioned adjacent to the translation initiating amino acid methionine. This modified hSODm permitted removal of the amino terminal methionine by processing enzymes.
A method of treating a patient having inflammatory joint disease or post-ischemic tissue injury is also provided.


REFERENCES:
patent: 82303600.9 (1983-01-01), None
patent: 85110560.1 (1986-02-01), None
patent: 0284105 (1988-02-01), None
patent: 8810480.5 (1988-09-01), None
patent: 0070656 (1983-01-01), None
patent: 0172577 (1986-02-01), None
patent: 2183658A (1987-10-01), None
Huhn-Edholm et al, cited in Chem. Abstracts, vol. 105:218641t (1986).*
Baret et al,cited in Chem. Abstracts,vol. 101:143782w (1984).*
Steinman, H.M., et al,J of Biol Chem(1974) 249 (22):7326-7338.
Jabusch, J.R., et al,Biochem(1980) 19:2310-2316.
Barra, D., et al,FEBS Letters(1980) 120(1):53-55.
Lieman-Hurwitz, J., et al,Proc Natl Acad Sci USA(1982) 79:2808-2811.
Barra, D., et al,J of Biol Chem(1984) 259 (20):12595-12601.
Tsunasawa, S., et al,J of Biol Chem(1985) 260(9):5382-5391.
Hallewell, R.A., et al,Nucl Acids Res(1985) 13(6):2017-2034.
Carlioz, A., et al,The EMBO Journal(1986) 5(3):623-630.
Barra et al., (1984) J. Biol. Chem 259(20):12595.
Carlioz et al., (1986) EMBO J. 5(3):623-630.
Tsunasawa et al., (1985) J. Biol. Chem. 260(9):5382:5391.
Hallewell et al., (1985) Nucleic Acids Research 13(6):2017-2034.
Jabusch et al., (1980) Biochemistry 19:2310-2316.
Barra et al., (1980) FEBS Letters 120(1):53-55.
Steinman et al., (1974) J. Biol. Chem 249(22):7326-7338.
Lieman-Hurwitz et al., (1982) Proc. Natl. Acad. Sci. USA 79:2808-2811.
Wilsman in Superoxide and Superoxide Dismutase in Chemistry, Biology, and Medicine, Proceedings of the 4th International Conference on Superoxide and Superoxide Dismutase held in Rome, Italy, Sep. 1-6, 1985 (Elsevier Publishers, edited by Guiseppe Rotilio), 1986, pp. 500-507.
Flohe et al., inDevelopments in Biochemistry, vol. 11B (Eds. E.M. Bannister and J.V. Bannister), Elsevier/North Holland, Amsterdam, 1980, pp. 424-430.

LandOfFree

Say what you really think

Search LandOfFree.com for the USA inventors and patents. Rate them and share your experience with other people.

Rating

Manganese superoxide dismutase cloning and expression in... does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Manganese superoxide dismutase cloning and expression in..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Manganese superoxide dismutase cloning and expression in... will most certainly appreciate the feedback.

Rate now

     

Profile ID: LFUS-PAI-O-2584696

  Search
All data on this website is collected from public sources. Our data reflects the most accurate information available at the time of publication.