Post-translational proteolytic processing of procollagen C-terminal proteinase enhancer releases a metalloproteinase inhibitor.
Activity of matrix metalloproteinases (MMP) is regulated by a family of proteins called tissue inhibitors of metalloproteinases (TIMP). Four TIMPs have been cloned, and their molecular weights range from 29,000 to 20,000. By reverse zymography, we have observed a metalloproteinase inhibitor with an apparent molecular weight of 16, 500 from medium conditioned by human brain tumor cells. Antibodies directed against TIMPs failed to react with the 16,500 molecular weight inhibitor, indicating that it was not a truncated form of a known TIMP. The inhibitor was isolated from conditioned medium using affinity and ion exchange chromatography. N-terminal sequences of the inhibitor matched amino acid sequences within the C-terminal domain of a protein known as procollagen C-terminal proteinase enhancer (PCPE). Thus, the inhibitor was named CT-PCPE. Comparison of the N-terminal domain of TIMP with CT-PCPE revealed that both contained six cysteine residues. As in the case of TIMP, reduction and alkylation abolished the inhibitory activity of CT-PCPE. Purified CT-PCPE inhibited MMP-2 with an IC(50) value much greater than that of TIMP-2. This implies that MMPs may not be the physiologic targets for CT-PCPE inhibition. However, these results suggest that CT-PCPE may constitute a new class of metalloproteinase inhibitor.
Mott
J D
JD
Department of Radiology, University of California, San Francisco, California 94143-0750, USA. JDMott@lbl.gov
Thomas
C L
CL
Rosenbach
M T
MT
Takahara
K
K
Greenspan
D S
DS
Banda
M J
MJ
D000595:Amino Acid Sequence; D001932:Brain Neoplasms; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D016326:Extracellular Matrix Proteins; D005341:Fibrinolysin / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D019714:Tissue Inhibitor of Metalloproteinases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; C491555:PCOLCE protein, human; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D005341:Fibrinolysin