Evidence for a triple helix recognition site in the hemopexin-like domains of human fibroblast and neutrophil interstitial collagenases.
Evidence for a triple helix recognition site in the hemopexin-like domains of human fibroblast and neutrophil interstitial collagenases.
复制标题
人成纤维细胞和中性粒细胞间质胶原酶的血红素结合蛋白样结构域中存在三螺旋识别位点的证据。
DOI：
10.1111/j.1749-6632.1994.tb24721.x
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发表时间：
1994
影响因子：
5.2
通讯作者：
VanWart,HE
中科院分区：
文献类型：
--
作者：
Netzel-Arnett,S;Salari,A;Goli,UB;VanWart,HE
材料和方法材料。在抑制实验之前，将Pro-HNC、pro-HFC、人成纤维细胞前基质溶解素（pro-HFS）和人前基质溶解素（pro-MAT）纯化至同质并活化。如前所述，Pro-HNC从人的皮毛中分离。1从人牙龈成纤维细胞收获培养基中纯化pro-HFC和pro-HFS。从携带扩增的pro-MAT cDNA的中国仓鼠卵巢细胞中分离Pro-MAT。12 [3 H]乙酰化底物根据Mallya等人13 Pro-OBzl.根据Bodanszky和B~ danszky的方法合成了TSA和Fmoc-6-氨基己酸（AHA）。DNP-Pro-Leu-Ala-Leu-Trp-Ala-Arg（DNP-PLALWAR）的合成和纯化方法与文献报道相同。化学品购自以下来源：Boc-Gly、Hyp-OBzl。HCl、Boc-Gly-PAM树脂、来自Bachem的Fmoc-Lys（E-BOC）;乙酸酐、二氯甲烷（DCM）、N，N-二甲基甲酰胺（DMF）、N-甲基吡咯烷酮（NMP）、二环己基碳二亚胺（DCC）、三氟甲磺酸（TFMSA）、异丙醇、乙醇、哌啶、三乙胺（TEA）、AHA、甲苯磺酸（TSA）、乙腈、二氧六环、醚、甲醇、三氯乙酸和三氟乙酸（TFA）
MATERIALS AND METHODSMaterials. Pro-HNC, pro-HFC, human fibroblast prostromelysin (pro-HFS), and human promatrilysin (pro-MAT) were purified to homogeneity and activated prior to the inhibition experiments. Pro-HNC was isolated from human buRy coats as described earlier. 1 Pro-HFC and pro-HFS were purified from the harvest media of human gingival fibroblasts. ll Pro-MAT was isolated from Chinese hamster ovary cells carrying an amplified pro-MAT cDNA. 12 [3H] acetylated substrates were prepared according to Mallya et al. l3 Pro-OBzl. TSA and Fmoc-6-aminohexanoic acid (AHA) were prepared according to Bodanszky and B~ danszky.~~ DNP-Pro-Leu-Ala-Leu-Trp-Ala-Arg (DNP-PLALWAR) was synthesized and purified as described previou~ ly.~~Chemicals were purchased from the following sources: Boc-Gly, Hyp-OBzl. HCI, Boc-Gly-PAM resin, Fmoc-Lys (E-BOC) from Bachem; acetic anhydride, dichloromethane (DCM), N, N-dimethylformamide (DMF), N-methylpyrrolidinone (NMP), dicyclohexylcarbodiimide (DCC), trifuoromethanesulfonic acid (TFMSA), isopropanol, ethanol, piperidine, triethylamine (TEA), AHA, toluene sulfonic acid (TSA), acetonitrile, dioxane, ether, methanol, trichloroacetic acid and trifluoroacetic acid (TFA)