BIOMAT Database Logo BIOMAT Database Logo

Preparation and reconstitution with divalent metal ions of class I and class II Clostridium histolyticum apocollagenases. 1988

E L Angleton, and H E Van Wart
Department of Chemistry, Florida State University, Tallahassee 32306.

Both gamma- and zeta-collagenases from Clostridium histolyticum are fully and reversibly inhibited by 1,10-phenanthroline at pH 7.5 in the presence of 10 mM CaCl2 with KI values of 0.11 and 0.040 mM, respectively. The inhibition is caused by removal of the single, active-site Zn(II) present in each of these enzymes. The nonchelating analogue 1,5-phenanthroline has no effect on the activity of either enzyme. Dialysis of the enzymes in the presence of 1,10-phenanthroline, followed by back dialysis against buffer containing no chelating agent, gives the respective apocollagenases. Both apoenzymes can be instantaneously and fully reactivated by the addition of 1 equiv of Zn(II). Variable amounts of activity are restored to both apocollagenases by Co(II) and Ni(II) and to gamma-apocollagenase by Cu(II). The activity titration curve for gamma-apocollagenase with Co(II) and Scatchard plots for the reconstitution of gamma-apocollagenase with Cu(II) and Ni(II) and of zeta-apocollagenase with Ni(II) and Co(II) indicate that all activity changes are the result of binding of a single equivalent of these divalent metal ions at the active site of the collagenases. Cd(II) and Hg(II) do not restore measurable activity to either apoenzyme.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008628 Mercury A silver metallic element that exists as a liquid at room temperature. It has the atomic symbol Hg (from hydrargyrum, liquid silver), atomic number 80, and atomic weight 200.59. Mercury is used in many industrial applications and its salts have been employed therapeutically as purgatives, antisyphilitics, disinfectants, and astringents. It can be absorbed through the skin and mucous membranes which leads to MERCURY POISONING. Because of its toxicity, the clinical use of mercury and mercurials is diminishing.
D008670 Metals Electropositive chemical elements characterized by ductility, malleability, luster, and conductance of heat and electricity. They can replace the hydrogen of an acid and form bases with hydroxyl radicals. (Grant & Hackh's Chemical Dictionary, 5th ed) Metal
D009532 Nickel A trace element with the atomic symbol Ni, atomic number 28, and atomic weight 58.69. It is a cofactor of the enzyme UREASE.
D010618 Phenanthrolines Phenanthroline
D002413 Cations, Divalent Positively charged atoms, radicals or groups of atoms with a valence of plus 2, which travel to the cathode or negative pole during electrolysis. Divalent Cations
D003012 Microbial Collagenase A metalloproteinase which degrades helical regions of native collagen to small fragments. Preferred cleavage is -Gly in the sequence -Pro-Xaa-Gly-Pro-. Six forms (or 2 classes) have been isolated from Clostridium histolyticum that are immunologically cross-reactive but possess different sequences and different specificities. Other variants have been isolated from Bacillus cereus, Empedobacter collagenolyticum, Pseudomonas marinoglutinosa, and species of Vibrio and Streptomyces. EC 3.4.24.3. Clostridiopeptidase A,Clostridium histolyticum Collagenase,Collagenase, Microbial,Collagenase Clostridium histolyticum,Collagenase-Like Peptidase,Collalysine,Nucleolysin,Clostridium histolyticum, Collagenase,Collagenase Like Peptidase,Collagenase, Clostridium histolyticum,Peptidase, Collagenase-Like,histolyticum, Collagenase Clostridium
D003013 Clostridium A genus of motile or nonmotile gram-positive bacteria of the family Clostridiaceae. Many species have been identified with some being pathogenic. They occur in water, soil, and in the intestinal tract of humans and lower animals.
D003035 Cobalt A trace element that is a component of vitamin B12. It has the atomic symbol Co, atomic number 27, and atomic weight 58.93. It is used in nuclear weapons, alloys, and pigments. Deficiency in animals leads to anemia; its excess in humans can lead to erythrocytosis. Cobalt-59,Cobalt 59
D003300 Copper A heavy metal trace element with the atomic symbol Cu, atomic number 29, and atomic weight 63.55. Copper-63,Copper 63

Related Publications

E L Angleton, and H E Van Wart
Effect of divalent metal ions on collagenase from Clostridium histolyticum.
June 1991, Biochemistry international,
E L Angleton, and H E Van Wart
Complementary substrate specificities of class I and class II collagenases from Clostridium histolyticum.
November 1985, Biochemistry,
E L Angleton, and H E Van Wart
Different roles of class I and class II Clostridium histolyticum collagenase in rat pancreatic islet isolation.
February 1995, Diabetes,
E L Angleton, and H E Van Wart
Kinetics of hydrolysis of type I, II, and III collagens by the class I and II Clostridium histolyticum collagenases.
February 1992, Journal of protein chemistry,
E L Angleton, and H E Van Wart
Isolation and characterization of collagenases I and II from Clostridium histolyticum.
September 1965, Biochimica et biophysica acta,
E L Angleton, and H E Van Wart
Limited proteolysis of type I collagen at hyperreactive sites by class I and II Clostridium histolyticum collagenases: complementary digestion patterns.
February 1987, Biochemistry,
E L Angleton, and H E Van Wart
Interactions of DNA with divalent metal ions. I. 31P-NMR studies.
January 1982, Biopolymers,
E L Angleton, and H E Van Wart
Interactions of divalent metal ions with inorganic and nucleoside phosphates. I. Thermodynamics.
December 1972, Journal of the American Chemical Society,
E L Angleton, and H E Van Wart
[Preparation of cavities for cast metal inlays in Black's Class I and Class II].
July 1951, L' Information dentaire,
E L Angleton, and H E Van Wart
Substrate recognition by the collagen-binding domain of Clostridium histolyticum class I collagenase.
March 2001, The Journal of biological chemistry,
Copied contents to your clipboard!