Expression,structure, and function of enamel proteinases

Proteinases serve two important functions during dental enamel formation: They (a) process and (b) degrade enamel proteins. Different enzymes carry out these functions. Enamelysin (MMP-20) is the foremost enamel matrix-processing enzyme. Its expression initiates prior to the onset of dentin mineralization and continues throughout the secretory stage of amelogenesis. In vitro, enamelysin catalyzes all of the amelogenin cleavages that are known to occur during the secretory stage in vivo, and it is probably the enzyme responsible for the processing of all enamel proteins. There is evidence suggesting that enamelysin activity is critical for proper enamel formation. Uncleaved and processed enamel proteins often segregate into different compartments within the developing enamel layer, suggesting that they may have different functions. Intact ameloblastin and its C-terminal cleavage products localize in the superficial rod and interrod enamel, while its N-terminal cleavage products congregate in the sheath space. Intact enamelin is only present at the mineralization front within a micrometer of the enamel surface, while its cleavage products concentrate in the rod and interrod enamel. Processed enamel proteins accumulate during the secretory stage, but disappear early in the maturation stage. Enamel matrix serine proteinase 1 (EMSP1), now officially designated kallikrein 4 (KLK4), is believed to be the predominant degradative enzyme that clears enamel proteins from the matrix during maturation. KLK4 expression initiates during the transition stage and continues throughout maturation. KLK4 concentrates at the enamel surface when the enamel matrix disappears, and aggressively degrades amelogenin in vitro. During tooth development, proteinases are secreted by ameloblasts into the extracellular space, where they cleave enamel proteins by catalyzing the hydrolysis of peptide bonds. Enamel proteinases are present in low abundance and are not likely to participate directly in the mineralization process. Two major enamel proteinases have been identified: enamelysin (MMP20) and kallikrein 4 (KLK4). These proteinases are expressed at different times and have different functions. Their roles are to modify and/or to eliminate enamel matrix proteins, which affects the way enamel proteins interact with each other and with the developing enamel crystallites. A brief review of dental enamel formation is presented, followed by a more detailed analysis of enamelysin and KLK4 expression, structure, and function.     

Structure and function of enamel gene products

The present paper reviews the main features of amelogenin and enamelin biochemistry, molecular biology, structural and ultrastructural localization, and immunology. It also examines recent studies concerning the origin, chemical characterization, suggested role, and participation of these two major classes of extracellular developing enamel matrix proteins in the complex process of "matrix-mediated" mineralization.     

The orientation of the enamel prisms at the enamel surface

The angulation that prisms obtain with the surface in human permanent teeth was analysed on broken enamel by means of the scanning electron microscope. In the cervical region the prisms end nearly perpendicular towards the surface, while the angle becomes more and more acute towards the occlusal region of the crown. In the cuspal area the deviation from the perpendicular direction of the prisms approaches 70 degrees. The size discrepancy between the inner and the outer surface of the enamel mantle can be explained by the more or less angulated position of the prisms.     

A novel structure in the stomach and intestine of two species of Australian marsupial mice

A tubular or sac-like structure with a well defined lumen which occasionally contained parasites was found in the outer muscle wall of the small and large intestine in 32 out of 40 male and 35 of 40 female marsupial mice of the two species, Antechinus swainsonii and Antechinus stuartii (Macleay). A similar structure was also found in the muscle layers of the stomach in 5 marsupial mice of both species. The cellular matrix of the structure appeared to be composed of randomly organized macrophage-like cells and in one case there was evidence of an inflammatory reaction to parasites within the lumen. The origin of this tube is either vestigial or the result of reactive changes.     

The ultrastructure of the enamel organ related to enamel formation

The ultrastructure of the cells of the enamel organ related to enamel formation was studied using the lower incisors of adult male rats. In the region of enamel deposition, stratum intermedium cells are stabilized by a system of intercellular bridges and intracellular fibrils. The mitochondria in these cells are positioned toward the extracellular channels through which any direct intercellular exchange between the capillaries and ameloblasts must occur. Tentatively, the mitochondrial arrangement is considered to be related to the movements of electrolytes and water across the capillary-ameloblast interval. In the region of transition, enamel deposition ceases and the ergastoplasm of the ameloblasts is removed, apparently by cytosegresomes, with an accompanying reduction in the height of the ameloblasts. Here, vesicles containing stippled material are infrequent compared to their occurrence in ameloblasts concerned with enamel deposition. Other vesicles, characteristically found in ameloblasts related to maturing enamel, first appear in the transition region and seem to originate from the cell membrane abutting on the enamel. In the region of maturation, cytosomes are common in the Golgi region whereas vesicles and mitochondria predominate in the distal ends of the ameloblasts. The papillary cells contain an unusually large number of mitochondria, elaborate microvilli and vesicles, which suggests that these cells are extremely active, presumably in the movement of materials related to enamel maturation. The changes in structure of the papillary cells, which occur concomitantly with those of the ameloblasts during enamel formation, are indicative of interrelated functional changes and strongly support the concept of ameloblasts and papillary cells acting together as a functional unit.     

Primary structure and variation of the T-cell receptor delta-chain from a marsupial,Macropus eugenii

Although gammadelta T-cells form only a small portion of circulating T-cells in mice and humans, they are more frequent in many other types of mammals and this has lead to speculation regarding their roles and the evolutionary significance of their relative abundance. Moreover, whilst clear homologues of four types of T-cell receptor (TCR) chains (alpha, beta, delta and gamma) have been identified in vertebrates as distantly related as eutherian mammals and cartilaginous fish, there are still many gaps in our knowledge of these TCR components from various taxa. Such knowledge would further illuminate the evolution and function of these receptors and of gammadelta T-cells. Here, we report the molecular cloning of a TCR-delta chain cDNA from the tammar wallaby (Macropus eugenii) which represents the first component of the gammadelta TCR to be characterised from a marsupial. A PCR-based survey of variable (V) segment usage in tammar wallaby mammary-associated lymph node indicated that, although gammadelta T-cells may be sparse in this type of tissue, this species has at least three subfamilies of V genes that have been broadly conserved across vertebrate evolution. Two V subfamilies found in the tammar wallaby were relatively similar and may have diverged more recently, an event that probably occurred at some point in the marsupial lineage.     

Helix structure of ribbon-like crystals in bovine enamel

In the early mineralized enamel crystals, ribbon-like crystals appear near the ameloblasts. Some ribbon-like crystals showed helical or spiral structure within restricted environment during the preparation of embryonic bovine specimens for electron microscope. These specimens did not suffer from the cutting damages nor staining effects. The main cause of the helix structure is considered a result of the dehydration during preparation. The periodic structure must reflect the regularity of initial enamel crystals. If dehydration caused the ribbon-like crystal to induce the periodic helix, it is one possibility that the earliest enamel crystal is OCP which has been proposed as a precursor of HA. Because it is considered that OCP is more sensitive to dehydration and more symmetric structure than biological HA. The periodicity of the helical ribbon-like structure was about 25 to 55 nm long and could be compared to the periodicity of organic helices which had observed in an immature rat enamel.     

The structure and function of the Niemann-Pick C1 protein

Niemann-Pick C (NPC) disease is a recessive cholesterol storage disorder characterized by severe, progressive neurodegeneration. The primary causative gene found on chromosome 18q11-12 was identified by a positional cloning approach. The NPC1 gene product is predicted to be a large polytopic glycoprotein with a cytoplasmic tail containing a dileucine endosome-targeting motif. The NPC1 protein sequence shares strong homology with a newly identified homologue, NPC1L1, and the morphogen receptor Patched. In addition, a group of five NPC1 transmembrane domains share homology with the sterol-sensing domain of proteins involved in cellular cholesterol homeostasis. Subcellular localization studies have shown NPC1 to reside in late endosomes and to transiently associate with lysosomes and the trans-Golgi network. Analysis of its topological arrangement in membranes suggests that NPC1 contains 13 transmembrane domains and three large, hydrophilic, lumenal loops. Currently, there is no direct evidence as to the function of the NPC1 protein; however, a number of observations suggest that NPC1 may be related to a family of prokaryotic efflux pumps and thus it may also act as a molecular pump.     

The development of enamel structure in rat incisors as compared to the teeth of monkey and man

The rat incisor is an excellent model system in which to study amelgenesis. However, the information obtained has not been extrapolated to the human because of alleged structural differences between the teeth. The obvious differences include continuous eruption in rat incisors and an enamel rod pattern in rats which seemingly differs from the keyhole pattern of human enamel. A comprehensive analysis was made of those features of enamel structure considered fundamental to the understanding of its formation. This was done by applying the knowledge of amelogenesis obtained in rat incisors to the teeth of monkey and man. The following points of basic similarity were established between these species: (1) Interrod enamel is secreted first. It forms the side walls of cavities which are initially occupied by Tomes' processes. (2) The formation of interrod cavities is followed by deposition of enamel rods within these spaces. (3) The rods conform to the shape of the cavities and are secreted from one surface of Tomes' process. (4) At the initial site of rod deposition its enamel is continuous with the interrod enamel wall. (5) Growth of the rod compresses the process to one side of the cavity resulting in an arcade-shaped "space" between the rod and the remaining interrod walls. This study demonstrates that it is no longer necessary to postulate a keyhole structure for primate enamel, and it has established that a fundamental similarity exists in the basic structure and in the mode of formation of enamel in all three species.     

CD14的结构和功能

ＣＤ１４分为膜上ＣＤ１４（ｍＣＤ１４）和可溶性Ｃ在４（ｓＣＤ１４）。ｍＣＤ１４２在单核细胞等表面：ＳＣＤ１４存在于正常人和动物的血清中,编码ＣＤ１４的基因已被克隆,该基因位于人５号常染色体长臂端５ｑ＾２３－ｑ＾３１,。ｍＣＤ１４是５５ｋＤａ糖蛋白,化学组成为糖基磷脂肌醇（ＧＰＩ）－蛋白质,通过ＰＩ的磷脂部分与细胞膜连拉;ｓＣＤ１４蛋白部分与ｍＣＤ１４基因相同,但不含ＰＩ部分,故分子量稍小,为４８ｋ     

The structure and function of interferon-gamma receptors.

Interferon-gamma exerts its pleiotropic immunomodulatory effects by interacting with a single type of IFN gamma receptor expressed on nearly all cells. Human and murine IFN gamma receptors have been purified, their cDNAs cloned and expressed, and their primary structure elucidated. In addition, a considerable amount of data is available that defines the life cycle of the receptor including the kinetics of its biosynthesis, its constitutive- and ligand-induced phosphorylation, and its recycling behavior. Recent transfection experiments have revealed that an additional species-specific component is necessary to form functionally active IFN gamma receptors in heterologous cells. On the basis of complementation assays, the gene for the human accessory molecule has been localized to human chromosome 21. However, the nature of the gene product(s) remains unknown. Using murine fibroblasts that contain a single copy of human chromosome 21 and eukaryotic cell expression vectors that contain a series of human IFN gamma receptor intracellular domain deletion mutants, we have been able to demonstrate that the receptor's intracellular domain plays an obligatory role in mediating IFN gamma-dependent biologic responses. Subsequent studies showed that two distinct regions of the intracellular domain are functionally important: a membrane proximal region of 48 amino acids needed for both internalization and induction of biologic responses and the carboxy terminal 39 amino acids needed only for function and not for internalization. Moreover, receptor-mediated ligand internalization was found to be insufficient for biologic response induction.(ABSTRACT TRUNCATED AT 250 WORDS)