Crystal structures of matrilysin-inhibitor complexes

The matrix metalloproteinases gelatinase A (MMP-2) and gelatinase B (MMP-9) are implicated in the physiological and pathological breakdown of several extracellular matrix proteins. In the present study, we show that long-chain fatty acids (e.g. oleic acid, elaidic acid, and cis-and trans-parinaric acids) inhibit gelatinase A as well as gelatinase B with K i values in the micromolar range but had only weak inhibitory effect on collagenase-1 (MMP-1), as assessed using synthetic or natural substrates. The inhibition of gelatinases depended on fatty acid chain length (with C18 > C16, C14, and C10), and the presence of unsaturations increased their inhibitory capacity on both types of gelatinase. Ex vivo experiments on human skin tissue sections have shown that micromolar concentrations of a long-chain unsaturated fatty acid (elaidic acid) protect collagen and elastin fibers against degradation by gelatinases A and B, respectively. In order to understand why gelatinases are more susceptible than collagenase-1 to inhibition by longchain fatty acids, the possible role of the fibronectin-like domain (a domain unique to gelatinases) in binding inhibitory fatty acids was investigated. Affinity and kinetic studies with a recombinant fibronectin-like domain of gelatinase A and with a recombinant mutant of gelatinase A from which this domain had been deleted pointed to an interaction of long-chain fatty acids with the fibronectin-like domain of the protease. Surface plasmon resonance studies on the interaction of longchain fatty acids with the three individual type II modules of the fibronectin-like domain of gelatinase A revealed that the first type II module is primarily responsible for binding these compounds. Matrix metalloproteinases (MMPs)1 compose a family of at least 23 related zinc-dependent endopeptidases (1) that are collectively able to degrade extracellular matrix proteins such as collagens, laminins, fibronectin, elastin, and proteoglycans. They are consequently implicated in physiological remodeling of connective tissue occurring in embryonic development and repair (2-4). Most of them are secreted as inactive proenzymes and are then extra-or pericellularly activated by other MMPs or serine proteinases (5). Their catalytic activities are strictly controlled by endogenous specific inhibitors designated as tissue inhibitors of metalloproteinases (TIMPs) (6) and also ␣ 2 -macroglobulin (7). The balance between activated MMPs and TIMPs determines the overall MMP proteolytic activity and consequently the extent of extracellular matrix degradation. Local disruption of the MMP-TIMP balance can lead to pathological degradative processes including rheumatoid arthritis, atherosclerosis, tumor growth, and metastasis (8 -10). MMPs are multidomain enzymes containing propeptide, catalytic and, except matrilysin (MMP-7), MMP-23, and endometase/matrilysin-2 (MMP-26), hemopexin-like domains. Gelatinase A (MMP-2) and gelatinase B (MMP-9) contain in addition three tandem copies of a 58-amino acid fibronectin type II (FN-II) modul...

Section: Effect Of Elaidic Acid On the Degradation Of Human Skin Collcontrasting

confidence: 54%

Involvement of Fibronectin Type II Repeats in the Efficient Inhibition of Gelatinases A and B by Long-chain Unsaturated Fatty Acids

Berton

Rigot

Huet

et al. 2001

“…These peptides were chosen based on knowledge gained from structure-function studies performed on other MMPs. The structures for the catalytic domains of several members of the MMP family have recently become available including human fibroblast collagenase (MMP-1) (22-24), human stromelysin-1 (MMP-3) (25,26), human matrilysin (MMP-7) (27), and human neutrophil collagenase (MMP-8) (28 -30). These structures revealed that the SЈ 1 subsite is the most well defined pocket in these MMPs and consists of a hydrophobic pocket which varies greatly in its depth.…”

Section: Figmentioning

confidence: 99%

“…These structures revealed that the SЈ 1 subsite is the most well defined pocket in these MMPs and consists of a hydrophobic pocket which varies greatly in its depth. Mutational analyses of the SЈ 1 pocket (29) revealed that residue 214 (numbering according to Browner (27)) which lies at the bottom of the SЈ 1 pocket is critical in determining its shape. For MMP-1 and MMP-7, which have an Arg 214 and a Tyr 214 , respectively, these residues point into the SЈ 1 pocket, thus forming a shallow pocket.…”

Section: Figmentioning

confidence: 99%

Hydrolysis of a Broad Spectrum of Extracellular Matrix Proteins by Human Macrophage Elastase

Gronski

Martin²,

Kobayashi

et al. 1997

277

204

Macrophage elastase (ME) was originally named when metal-dependent elastolytic activity was detected in conditioned media of murine macrophages. Subsequent cDNA cloning of the mouse and human enzyme demonstrated that ME is a distinct member of the matrix metalloproteinase family. To date, the catalytic parameters that describe the hydrolysis of elastin by ME have not been quantified and its activity against other matrix proteins have not been described. In this report, we have examined the action of purified recombinant human ME (rHME), produced in Escherichia coli, on elastin and other extracellular matrix proteins. On a molar basis, rHME is approximately 30% as active as human leukocyte elastase in solubilizing elastin. rHME also efficiently degrades ␣ 1 -antitrypsin (␣ 1 -AT), the primary physiological inhibitor of human leukocyte elastase. In addition, rHME efficiently degrades fibronectin, laminin, entactin, type IV collagen, chondroitan sulfate, and heparan sulfate. These results suggest that HME may be required for macrophages to penetrate basement membranes and remodel injured tissue during inflammation. Moreover, abnormal expression of HME may contribute to destructive processes such as pulmonary emphysema and vascular aneurysm formation. To further understand the specificity of HME, the initial cleavage sites in ␣ 1 -AT have been determined. In addition, the hydrolysis of a series of synthetic peptides with different P 1 residues has been determined. rHME can accept large and small amino acids at the P 1 site, but has a preference for leucine.Macrophage elastase (MMP-12) shares many properties with other members of the matrix metalloproteinase (MMP) 1 gene family, yet it is unique in several ways. Like other MMPs, metalloelastase requires zinc for catalytic activity, is inhibited by the tissue inhibitors of metalloproteinases (TIMPs), and has common structural domains with other MMPs (1, 2). Human macrophage elastase (HME) is most closely related to collagenase-1 (MMP-1) and stromelysin-1 (MMP-3), being 49% identical to each at the amino acid level (3). Moreover, the gene for macrophage elastase, composed of a common 10-exon, 9-intron structure, is on human chromosome 11q22.2/22.3 with at least six other MMPs (4). Despite these similarities, HME possesses certain distinct biological and biochemical properties. Expression appears to be largely restricted to tissue macrophages (4). Upon activation, it not only cleaves its 8-kDa N-terminal domain, but also has a unique propensity to autolytically release its 23-kDa C-terminal domain resulting in a mature active 22-kDa proteinase (3)(4)(5).Macrophage elastase shares its elastolytic activity (6, 7) with only a few MMPs, including the gelatinases (MMP-2 and MMP-9) and, to a lesser extent, matrilysin (8, 9). However, despite this characteristic activity, the relative capacity of metalloelastase (human or mouse) to degrade elastin has never been quantified. In addition, the catalytic capacity of metalloelastase against other extracellular matrix components has...

“…ics, hydroxamic acid derivatives (2,14,21,22), phosphinamides and sulfodiimines (23,24), thiadiazole (25), and malonic acid derivatives (26,27).…”

mentioning

confidence: 99%

The 1.8-Å Crystal Structure of a Matrix Metalloproteinase 8-Barbiturate Inhibitor Complex Reveals a Previously Unobserved Mechanism for Collagenase Substrate Recognition

Brandstetter

Grams

Glitz

et al. 2001

124

103

The individual zinc endoproteinases of the tissue degrading matrix metalloproteinase (MMP) family share a common catalytic architecture but are differentiated with respect to substrate specificity, localization, and activation. Variation in domain structure and more subtle structural differences control their characteristic specificity profiles for substrates from among four distinct classes (Nagase, H., and Woessner, J.