C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/15 (2006.01) A61K 38/57 (2006.01) A61K 39/395 (2006.01) C07K 7/08 (2006.01) C07K 14/81 (2006.01) C07K 16/38 (2006.01) C12Q 1/68 (2006.01) G01N 33/573 (2006.01)
Patent
CA 2046649
The present invention is an isolated protein of 21,600 daltons which binds to both latent and activated type IV collagenase with high affinity at 1:1 molar stoichiometry, thereby abolishing enzyme activity. The protein is purified by affinity chromatography on solid phase metalloproteinase, or solid phase metalloproteinase substrates which bind the enzyme-inhibitor complex. The complete primary structure of this protein (initially called CSC-21K), as determined by sequencing overlapping peptides spanning the entire protein, reveals homology with a protein called TIMP, Tissue Inhibitor of Metalloproteinases. In addition, a cDNA for this novel inhibitor, now designated TIMP-2, was cloned from a melanoma cell and its sequence was compared with that of human TIMP-1. Northern blots of melanoma cell mRNA showed two distinct transcripts of 0.9 kb and 3.5 kb which are down-regulated by transforming growth factor-beta, and are unchanged by phorbol ester treatment. The figure 7 shows the cDNA and corresponding protein sequence of TIMP-2.
Krutzsh Henry
Liotta Lance A.
Stetler-Stevenson William G.
Department Of Health And Human Services The United States Of America As Represented By The Secretary
Gowling Lafleur Henderson Llp
U.s. Departm Ent Of Commerce The United States Of America Represented By The Secretary
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