title: The influence of flavonoid compounds on the in vitro inhibition study of a human fibroblast collagenase catalytic domain expressed in E. coli.
abstract: The human fibroblast collagenase catalytic domain (MMP1ca) that is considered a prototype for all interstitial collagenase and plays an important role in the turnover of collagen fibrils in the matrix was expressed as an inclusion body in the Escherichia coli. The purified enzyme displayed activity with substrate Dnp-Pro-Leu-Ala-Leu-Trp-Ala-Arg-OH with a K(m) value of 26.61±1.42 μM. The inhibition activity of the nine flavonoid compounds and gallic acid against MMP1ca was examined. Among the compounds tested, the IC(50) of seven flavonoid compounds were determined and ranged from 14.13 to 339.21 μM. Epigallocatechin gallate (EGCG) showed the highest inhibition toward MMP1ca with IC(50) values of 14.13±0.49 μM. EGCG showed a competitive inhibition pattern with a K(i) value of 10.47±0.51 μM. The free binding energy of EGCG against MMP1ca was -13.07 kcal mol(-1), which was calculated by using Autodock 3.0.5 software and showed numerous hydrophobic and hydrogen bond interactions. The galloyl group of EGCG, gallocatechin gallate and epicatechin gallate was determined to be important for inhibitory activity against MMP1ca.
authors.affiliation: School of Biological Sciences and Technology, Chonnam National University, 77 Yongbong-ro, Buk-gu, Gwangju 500-757, Republic of Korea.
authors.forename: Thi Thanh Hanh
authors.initials: TT
authors.lastname: Nguyen
authors.forename: Young-Hwan
authors.initials: YH
authors.lastname: Moon
authors.forename: Young-Bae
authors.initials: YB
authors.lastname: Ryu
authors.forename: Young-Min
authors.initials: YM
authors.lastname: Kim
authors.forename: Seung-Hee
authors.initials: SH
authors.lastname: Nam
authors.forename: Mi-Sook
authors.initials: MS
authors.lastname: Kim
authors.forename: Atsuo
authors.initials: A
authors.lastname: Kimura
authors.forename: Doman
authors.initials: D
authors.lastname: Kim
mesh_terms: D001667:Binding, Competitive; D002384:Catalysis / Q000187:drug effects; D020134:Catalytic Domain; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D017364:Collagenases / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli; D005347:Fibroblasts / Q000201:enzymology*; D005419:Flavonoids / Q000737:chemistry / Q000494:pharmacology*; D005707:Gallic Acid / Q000494:pharmacology; D006801:Humans; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D066298:In Vitro Techniques; D020128:Inhibitory Concentration 50; D020781:Matrix Metalloproteinase 1 / Q000187:drug effects* / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology*; D008958:Models, Molecular; D015394:Molecular Structure; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D005419:Flavonoids; D061965:Matrix Metalloproteinase Inhibitors; D011993:Recombinant Fusion Proteins; D005707:Gallic Acid; D002392:Catechin; C045651:epigallocatechin gallate; D017364:Collagenases; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1