Abstract:
:Ameloblastic tissue samples from unerupted bone molars were used to prepare subcellular enamel protein kinase preparations, nuclear + plasma membrane, cytosolic and microsomal, and used in in vitro phosphorylation of purified 20 kDa bovine amelogenin in the presence of 32P-ATP. Both cytosolic and microsomal preparations can phosphorylate purified native amelogenins, the addition of Ca2+ slightly increased the microsomal enzyme activity or at least did not inhibit the activity, whereas the presence of Ca2+ substantially decreased the cytosolic kinase activity towards phosphorylation of amelogenins. A comparative analysis using the enamel microsomal kinase against osteopontin, dephosphorylated casein and bone sialoprotein showed no phosphorylation of the first two proteins, and only minor phosphorylation of the bone sialoprotein. Overall, the present work demonstrates for the first time that the protein kinase responsible for the phosphorylation of amelogenins is a novel kinase, which is not inhibited by Ca2+, unlike the microsomal protein kinase (casein kinase type-II) of bone which phosphorylates secretory proteins osteopontin and bone sialoprotein and is strongly CaZ+ inhibited. The direct phosphoserine analysis on the purified bovine 20 kDa amelogenin indicated the presence of 0.8 moles of phosphoserine/mole protein naturally occurring, consistent with the quantitative analysis of 14C-radiolabeling of phosphoserines by conversion to dehydroalanine and in situ reaction with the thiol agent, 14C-mercaptoethanol, 0.64 moles 14C-incorporated/mole 20 kDa amelogenin. The purified low Mramelogenins 5.3 kDa E4 (TRAP) and 7.2 kDa E3 (LRAP), were also derivatized by 14C-mercaptoethanol, providing 0.46 and 0.88 moles 14C-incorporated/mole respectively. Further studies of the 14C-radiolabeled E4 amelogenin by sequence analysis confirmed one site of label to be at position 16 from the N-terminal and hence provided a direct evidence for the naturally occurring phosphoserine residue at this position.
journal_name
Connect Tissue Resjournal_title
Connective tissue researchauthors
Salih E,Huang JC,Strawich E,Gouverneur M,Glimcher MJdoi
10.3109/03008209809017041keywords:
subject
Has Abstractpub_date
1998-01-01 00:00:00pages
225-35; discussion 241-6issue
1-4eissn
0300-8207issn
1607-8438journal_volume
38pub_type
杂志文章abstract::Abnormal polyanion metabolism in cultured fibroblasts from patients with pseudoxanthoma elasticum (PXE) was studied by incubating normal and PXE fibroblasts in culture medium containing 35SO4 for 96 hr and measuring differences in secreted 35SO4-labelled proteoglycans (35S-PG). PXE medium contained less high molecular...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207809152287
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abstract::The purpose of this investigation is to evaluate enamel thickness in extant and extinct hominoids. The material used in this study spans the evolutionary history of this group, from 20 million years ago to the present. The objectives of this investigation are to test three hypotheses: (1) the Loading Hypothesis: loadi...
journal_title:Connective tissue research
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abstract::Cells derived from organ-explant culture of the cartilaginous component of osteochondrophytic spurs of human femoral heads were incubated with [35S]-sulfate in order to study sulfated proteoglycan biosynthesis in vitro. Secondary monolayer cultures incorporated [35S]-sulfate into macromolecules which were recovered in...
journal_title:Connective tissue research
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abstract:BACKGROUND:The presence of genetic variants in uric acid (UA) transporters can be associated with hyperuricemia, and therefore with an increased risk of monosodium urate (MSU) crystal precipitation. The inflammatory process triggered by these crystals leads to cartilage damage, which, in turn, could promote knee osteoa...
journal_title:Connective tissue research
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abstract::The mechanical properties of the human supraspinatus tendon (SST) are highly heterogeneous and may reflect an important adaptive response to its complex, multiaxial loading environment. However, these functional properties are associated with a location-dependent structure and composition that have not been fully eluc...
journal_title:Connective tissue research
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abstract::Studies were designed to test the hypothesis that plasma- and serum-deprived embryonic cells and tissues in vitro are capable of producing growth regulating factors which augment cartilage, bone and tooth induction during mouse mandibular process development. Embryonic mouse first branchial arch-derived mandibular pro...
journal_title:Connective tissue research
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abstract::The connective tissue protein elastin is largely responsible for maintaining the elasticity of major blood vessels and lung tissue. Comparatively few studies have been made of the immunologic properties of elastin because of its high degree of insolubility and because of its apparent limited capacity to elicit precipi...
journal_title:Connective tissue research
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doi:10.3109/03008208109152131
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abstract::Odontogenesis involves a complex series of processes including epithelial-mesenchymal interactions, morphogenesis, differentiation, fibrillogenesis, and mineralization. Extracellular (ECM) remodeling plays a critical role in the rapid morphological changes that accompany these events. It is proposed that matrix metall...
journal_title:Connective tissue research
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abstract::Gene expression of osteopontin (OPN) has been investigated in mice by application of laser capture microdissection (LCM) and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. LCM permits individual cells to be isolated ("captured") from tissue sections for molecular analyses. In this study, chondrocyt...
journal_title:Connective tissue research
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doi:
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abstract::The total and individual glycosaminoglycan (GAG) content at various stages of the Dupuytren disease process and in samples of normal palmar connective tissue (palmar dermis, palmar fascia and digital flexor tendon) from the hands of uninvolved age-matched controls have been assayed and compared. Morphological comparis...
journal_title:Connective tissue research
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更新日期:1982-01-01 00:00:00
abstract:OVERVIEW:Periostin (POSTN) is critical to bone and dental tissue morphogenesis, postnatal development, and maintenance; however, its roles in tissue repair and regeneration mediated by human periodontal ligament mesenchymal stem cells (PDLSCs) remain unclear. The present study was designed to evaluate the effects of PO...
journal_title:Connective tissue research
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abstract::Molecular distributions of the intermolecular cross-links in fetal bovine bone type I collagen fibrils were quantitatively determined and compared with those of periodontal ligament. Results indicated that Hyl and Lys residues in the COOH-terminal nonhelical peptide portions (residues 16C) of both alpha 1 chains were ...
journal_title:Connective tissue research
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更新日期:1989-01-01 00:00:00
abstract::Purpose: Complications including arthrofibrosis have been reported after anterior cruciate ligament reconstruction (ACLR) even under accelerated rehabilitation. To overcome this, we developed an animal model of ACLR-induced arthrofibrosis without immobilization.Materials and Methods: Thirteen male Wistar rats were div...
journal_title:Connective tissue research
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abstract::Quantitative levels of mRNAs coding for elastin, types I and III procollagen and gamma-actin were measured in porcine vascular material following balloon catheterization. A balloon catheter was introduced into the thoracic aorta and jugular vein of 3-6 week old pigs; following distention and six days of postoperative ...
journal_title:Connective tissue research
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doi:10.3109/03008208809008059
更新日期:1988-01-01 00:00:00
abstract::Anterior cruciate ligament (ACL) has poor healing ability and an injured ACL would induce the degeneration of other intra-articular connective tissues. However, the coordinated expression and activities of matrix metalloproteinase (MMPs) in intra-articular tissues induced by ACL rupture were poorly understood. With a ...
journal_title:Connective tissue research
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更新日期:2009-01-01 00:00:00
abstract::STR/ort mice develop a naturally occurring osteoarthritis (OA) of the knee joints. However, the evaluation of early OA changes has been difficult due to variability caused by gender, individual differences, and differences between the right and left lower limbs. The objective of this study was to analyze the variabili...
journal_title:Connective tissue research
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更新日期:2009-01-01 00:00:00
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journal_title:Connective tissue research
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更新日期:2001-10-01 00:00:00
abstract::Aggrecan is the prominent proteoglycan in cartilage and is modified with approximately 100 chondroitin sulfate (CS) chains through a tetrasaccharide linkage structure. In osteoarthritis (OA), the viscoelastic properties of cartilage are compromised on both the quantity and integrity of aggrecan core protein expressed ...
journal_title:Connective tissue research
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doi:10.3109/03008207.2012.756871
更新日期:2013-01-01 00:00:00
abstract::Transforming growth factor beta (TGF-beta) is a ubiquitous regulator of cellular growth and differentiation. The present study investigated the effects of TGF-beta on chick growth plate chondrocyte proliferation, matrix synthesis, and alkaline phosphatase activity in short term cultures. TGF-beta markedly stimulated D...
journal_title:Connective tissue research
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doi:10.3109/03008208909023900
更新日期:1989-01-01 00:00:00
abstract::Alveolar mucosa and attached gingiva are two continuous but functionally distinct connective tissues covering alveolar bone of the jaw. In this study, the major matrix component of these tissues, collagen, was biochemically characterized and compared. The tissues were obtained from mature pigs and analyzed for collage...
journal_title:Connective tissue research
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abstract::The matrix metalloproteinases collagenase (MMP-1) and stromelysin (MMP-3) are often coordinately expressed, and their promoters contain similar regulatory elements, including an AP-1 site at about -70. There are, however, additional sequences including an adjacent ETS site at about -90 in both promoters, and a NIP (nu...
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abstract:PURPOSE:Experimental measurements of bone mineral density distributions (BMDDs) enable a determination of secondary mineralization kinetics in bone, but the maximum degree of mineralization and how this maximum is approached remain uncertain. We thus test computationally different hypotheses on late stages of bone mine...
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abstract::DNA synthesis in rat and rabbit polyvinyl sponge induced granulation tissue has been studied using thymidine (methyl-3H). Synthesis was determined by measurement of thymidine incorporation into cold trichloroacetic acid insoluble material and by autoradiography. Granulation tissue was removed and immediately incubated...
journal_title:Connective tissue research
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更新日期:1976-01-01 00:00:00
abstract::Objective: Aseptic loosening is a major problem in total joint replacement. Implant wear debris provokes a foreign body host response and activates cells to produce a variety of mediators and ROS, leading to periprosthetic osteolysis. Elevated ROS levels can harm proteasome function. Proteasome inhibitors have been re...
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abstract::Two overlapping clones encompassing 8.5 kb of the human elastin gene were isolated from two genomic libraries constructed by partial digestion with either HaeIII/AluI or Sau3A and contained in lambda Charon 4A or EMBL3, respectively. The 6 kb of DNA comprising the most 3' portion of the gene were sequenced demonstrati...
journal_title:Connective tissue research
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更新日期:1987-01-01 00:00:00
abstract::Primary chondrocytes cultured in agarose can escape the gel, accumulate at the interface between agarose and the culture medium, and form an outgrowing tissue. These outgrowths can appear as voluminous cartilage-like nodules that have never been previously investigated. In the present study, bovine articular chondrocy...
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abstract::Rat bone marrow cells were cultured in vitro in a collagen-gel medium at 0.5% fetal bovine serum concentration for 10 days in the presence of recombinant human transforming growth factor-beta-1, genetically engineered to contain a collagen binding domain (rhTGF-beta1-F2), or a commercial rhTGF-beta1. To compare the ef...
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更新日期:2003-01-01 00:00:00
abstract::Purpose: Non-pathological child cortical bone (NPCCB) studies can provide clinicians with vital information and insights. However, assessing the anisotropic elastic properties of NPCCB remains a challenge for the biomechanical engineering community. For the first time, this paper provides elastic moduli values for NPC...
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