RESUMEN
The extracellular matrix protein 1 (ECM1) is an 85 kDa secreted glycoprotein, comprising four variants and playing a pivotal role in endochondral bone formation, angiogenesis, and tumour biology. A computational model for the three-dimensional structure of ECM1a was determined to identify the potential and/or concealed region(s) for binding with candidate partners in human skin. Multiple alignments for the secondary structure of ECM1a and b revealed similarity with serum albumin. The N-terminal domain of ECM1a consists mainly of alpha-helices (alphaD1), while the remaining three domains, namely serum albumin subdomain-like (SASDL) domains 2-4, were topologically comparable with the subdomain of the third serum albumin domain. Yeast-two-hybrid screening of a human foreskin cDNA library using both full-length ECM1a and the hot spot region for ECM1 gene mutations in lipoid proteinosis, an autosomal recessive genodermatosis (complete SASDL2 and the linker to SASDL3: aa177-aa361), as bait, isolated seven extracellular proteins. The site-specific interaction of ECM1a with two of these candidate binders, laminin 332 beta-3 chain and fibulin-3, was confirmed by in vitro and in vivo co-immunoprecipitation experiments. Immunohistologically both binders co-localized with ECM1 in human skin. Together, ECM1 is a multifunctional binding core and/or a scaffolding protein interacting with a variety of extracellular and structural proteins, contributing to the maintenance of skin integrity and homeostasis. Hence, disruption of the ECM1 function may cause the failure of multi-communication among the surrounding skin interstitial molecules, as seen in lipoid proteinosis pathology.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Dominios y Motivos de Interacción de Proteínas/fisiología , Secuencia de Aminoácidos , Sistema Libre de Células/metabolismo , Células Cultivadas , Epidermis/metabolismo , Proteínas de la Matriz Extracelular/genética , Fibroblastos/metabolismo , Humanos , Inmunoprecipitación , Queratinocitos/metabolismo , Datos de Secuencia Molecular , Unión Proteica/fisiología , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Albúmina Sérica/genética , Piel/metabolismo , Técnicas del Sistema de Dos Híbridos , Proteína de Unión a Vitamina D/genética , KalininaRESUMEN
The extracellular matrix protein 1 (ECM1) is expressed around blood vessels, which suggest a role for ECM1 in angiogenesis. Recombinant ECM1 stimulates proliferation of cultured endothelial cells and promotes blood vessel formation in the chorioallantoic membrane of chicken embryos. These observations make ECM1 a possible trigger for angiogenesis, tumor progression and malignancies. Interaction of ECM1 with perlecan, MMP-9 and fibulin-1C/D contributes to this hypothesis. However, the importance of ECM1 in cancer biology has been neglected so far. Nevertheless, a survey of ECM1 expression in different tumors indicated that ECM1, although not tumor specific, is significantly elevated in many malignant epithelial tumors that give rise to metastases, emphasizing its relevance in the cancer process.
Asunto(s)
Proteínas de la Matriz Extracelular/fisiología , Proteínas de Neoplasias/fisiología , Neoplasias/fisiopatología , Proteínas de Unión al Calcio/fisiología , Transformación Celular Neoplásica , Progresión de la Enfermedad , Femenino , Proteoglicanos de Heparán Sulfato/fisiología , Humanos , Masculino , Metaloproteinasa 9 de la Matriz/fisiología , Modelos Biológicos , Neoplasias/irrigación sanguínea , Neoplasias/patología , Neovascularización Patológica/fisiopatología , Especificidad de Órganos , Fragmentos de Péptidos/fisiología , Transducción de Señal , beta Catenina/fisiologíaRESUMEN
BACKGROUND: Extracellular matrix protein 1 (ECM1) is a secreted protein expressed in skin. Its dermatological relevance has been highlighted by the discovery of loss-of-function mutations in ECM1 in patients with lipoid proteinosis (LiP). OBJECTIVES: To determine the role of ECM1 in epidermal differentiation by examining gene and protein expression of epidermal differentiation markers in individuals with LiP and histological assessment of transgenic mouse skin that overexpresses Ecm1a in basal or suprabasal epidermis. METHODS: Subconfluent, confluent and postconfluent LiP and control keratinocyte cultures were analysed by Northern and Western blotting for differences in expression of differentiation markers. Expression of these markers was analysed in skin of patients with LiP by immunohistochemistry. To study effects of Ecm1 overexpression on epidermal differentiation, transgenic mice were generated under control of either a keratin 14 or an involucrin promoter. RESULTS: No differential expression of the different markers analysed was observed in LiP keratinocytes compared with controls. No histological differences were found in Ecm1-overexpressing mouse skin compared with wild-type. CONCLUSIONS: Absence of ECM1 does not lead to differences in epidermal differentiation. Moreover, overexpression of Ecm1a in vivo does not exert dramatic effects on epidermal structure. Collectively, these findings suggest no role of ECM1 in epidermal differentiation.
Asunto(s)
Epidermis/patología , Proteínas de la Matriz Extracelular/fisiología , Proteinosis Lipoidea de Urbach y Wiethe/patología , Adulto , Animales , Diferenciación Celular , Células Cultivadas , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/metabolismo , Humanos , Queratinocitos/metabolismo , Proteinosis Lipoidea de Urbach y Wiethe/metabolismo , Ratones , Ratones Transgénicos , Mutación , Piel/metabolismo , Piel/patologíaRESUMEN
BACKGROUND: The extracellular matrix protein 1 (ECM1) is expressed in human skin and plays an important role in its normal structure and function. In the rare genetic skin disease lipoid proteinosis, which is characterized by a loss-of-function mutation in the ECM1 gene, skin areas habitually exposed to the sun may show a more severely scarred and photoaged appearance. However, no data are available on the possible involvement of ECM1 expression in intrinsic and extrinsic skin ageing. OBJECTIVES: We hypothesized that ECM1 expression in human skin is regulated by age- and ultraviolet (UV)-dependent mechanisms. METHODS: Skin biopsies from 12 patients with histologically confirmed solar elastosis, from non-UV-exposed sites of 12 age-matched controls and 12 young subjects were analysed. To evaluate the influence of acute UV exposure, buttock skin of 10 healthy subjects was irradiated repetitively on 10 days with a solar simulator and compared intraindividually with non-UV-treated contralateral sites. The expression of ECM1 was investigated by immunohistochemistry using an ECM1 antibody detecting ECM1a and ECM1c isoforms. Semiquantitative analysis of staining intensity was carried out by densitometric image analysis. RESULTS: In normal human skin ECM1a and ECM1c are expressed mainly in the basal cell layers of epidermal keratinocytes and in dermal vessels. For the first time, an expression in the outer root sheath of hair follicles, in sebaceous lobules and epithelium of sweat glands is described. Intrinsically (UV-protected) aged skin shows a significantly reduced expression in basal and upper epidermal cell layers compared with young skin. In photoaged skin, expression is significantly increased within the lower and upper epidermis compared with age-matched UV-protected sites. Importantly, after acute UV exposure in young healthy subjects expression of ECM1 is markedly increased in both lower and upper epidermal cell layers. CONCLUSIONS: This is the first study to demonstrate a regulation of ECM1 expression in human skin by age and UV exposure. These data suggest that ECM1 expression may represent a cutaneous stress response to acute and chronic UV irradiation.