Collagenase-inhibitor complex

Brandstetter, H., Grams, F., Glitz, D., et al. (2001) The 1.8-A crystal structure of a matrix metalloproteinase 8-barbiturate inhibitor complex reveals a previously unobserved mechanism for collagenase substrate Recognition Journal of Biological Chemistry, 276, 17405-17412 corrigendum ibid., 276, 31474. 

Mouse and chick bone explants have been shown to produce a latent collagenase.33,34 jn addition, the mouse bone explants produce a latent neutral proteinase. Limited proteolysis of these molecules gives rise to proteoglycan and collagen degrading action. Latency may be due to the presence of an enzyme inhibitor complex. The chick latent collagenase has an apparent MW of 54,000, while the active form has an MW of 43,000. 3 Trypsin activation of both latent mouse enzymes decreases their MW s from 60-70,000 to about 40,000. The mouse neutral protease is inhibited by EDTA, cysteine, and serum.34... 

An advantage of NMR spectroscopy is the analysis of protein dynamics. Measurement and analysis of the relaxation parameters R1 R2, and the 15N NOE of 15N-labeled proteins leads to an order parameter (S2) that can describe the relative mobility of the backbone of the protein. Both collagenase-1 and stromelysin-1 have been studied either as inhibited complexes or the free protein [19, 52], Stromleysin-1 was studied with inhibitors binding to prime or nonprime subsites. Presence or absence of inhibitors in the nonprime sites had minor effects on the highly ordered structure of residues in these subsites, which are in contact with the... 

Bartoki N, Winkler FK, Williams DH, D Arcy A, Broadhurst MJ, Brown PA, Johnson WH, Murray EJ. Structure of the catalytic domain of human fibroblast collagenase complexed with an inhibitor. Nat Struct Biol 1994 1 106-110. 

Lovejoy B, Cleasby A, Hassell AM, Longley K, Luther MA, Weigl D, McGeehan G, McElroy AB, Drewry D, Lambert MH, Jordan SR. Structure of the catalytic domain of fibroblast collagenase complexed with an inhibitor. Science 1994 263 375-377. 

Fig. 30. X-ray crystal structure of batimastat-human neutrophil collagenase complex, showing the inhibitor coordinated to the catalytic Zn(II) (PDBID 1JAQ). Adapted from (576).

Grams F, Crimmin M, Hinnes L, Huxley P, Pieper M, Tschesche H, Bode W. Structure determination and analysis of human neutrophil collagenase complexed with a hydroxamate inhibitor. Biochemistry 1995 34 14012-14020. 

Eco is a remarkable inhibitor. It binds to and inhibits proteases with a broad range of pi values from 3.92 (collagenase) to 9.5 (grB) forming an E2P2 complex that typically buries between 1900 and 2500 A. This large and strongly associated complex is able to form crystals at a range of pH values from pH 5.0 to pH 8.8 (Tab. 7.1). To bind serine proteases efficiently with many different substrate prefer-... 

The glyoxylate-ene reaction, promoted by the Ti-BINOL complex, produces chiral a-hydroxy esters, which provide an easy access to the corresponding carboxylic acid derivatives bearing a chiral center at the a position. The adduct between the glyoxylate and exo olefin is proposed as a key intermediate of the collagenase-selective inhibitor, Trocade (Hoffmann-La Roche) [20]. This remarkable process has been developed for large-scale production. 

Figure 10. Comparison of the details of inhibitor binding in the two families superposition of the catalytic centers of collagenase (Fig. 6) and thermolysin (Fig. 9). The catalytic zinc atom is shown as a cyan sphere, with the collagenase-in-hibitor complex in yellow and the thermolysin complex shown in magenta. Thin cylinders denote putative hydrogen bonds. The green sphere shows the location of a structural calcium atom observed in collagenase. For clarity, only the alpha-carbon atoms of the proteins are shown. The differences in the relative oreintation of the subsites are evident.

In an analogous manner, inhibitors bearing substituents at the carbon spacer between the zinc ligand and the alpha centre of the PI subsite have different enzyme-inhibitor interactions with separate members of the MMP family. The crystal structure of the synthetic inhibitor RO32-0554 (5) (Fig. 13) complexed with fibroblast collagenase indicates a favorable interaction between the imide oxygen of the naphthalimide group of the inhibitor and an... 

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