• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.28 no.1
• /
• pp.31-41
• /
• 2002

Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

• Journal of fish pathology
• /
• v.25 no.1
• /
• pp.11-20
• /
• 2012

C-type lectins are crucial for pathogen recognition, innate immunity, and cell-cell interactions. In this study, a C-type lectin gene was cloned from the rock bream. The full-length RbCTL cDNA was 729 bp with a 429 bp ORF encoding a 164-residue protein. The deduced amino acid sequence of RbCTL had all of the conserved features crucial for its fundamental structure, including the four cysteine residues involved in sulfide bridge formation and potential $Ca^2+$/carbohydrate-binding sites. RbCTL contains a signal peptide one single carbohydrate recognition domain. It showed 29.4% similarity to the C-type lectin of rainbow trout. RbCTL mRNA was predominately expressed in gill and head-kidney tissue and expressed less in peripheral blood leukocytes, trunk-kidney, spleen, liver, intestine and muscle. Expression of RbCTL was differentially upregulated in rock bream stimulated with LPS, Con A/PMA and poly I:C.

• The Journal of Korean Academy of Prosthodontics
• /
• v.42 no.3
• /
• pp.301-306
• /
• 2004

Statement of problem. Fibronectin mediates its biological effects by binding to integrins on cell membranes through a consensus site including the Arg-Gly-Asp (RGD) sequence within tenth type III module. Purpose. The purpose of our study was to investigate the adsorption affinity of human recombinant fibronectin peptide (hFNIII 9-10) to titanium and to investigate the effect of the surrounding ionic composition on the adsorption process. Material and methods. As for evaluating the affinity of hFNIII 9-10 to Ti, titanium disks were incubated in 40, 80 and $120{\mu}g/ml$ hFNIII 9-10 solution at $37^{\circ}C$ overnight, repectively. As for evaluating the effect of surrounding ionic concentration, hFNIII 9-10 was dissolved in distilled water, phosphate buffered saline and RPMI 1640. Optical density (O.D.) was measured in ELISA reader. Results. The results were as follows; 1. The adsorption of hFNIII 9-10 showed significantly highest mean optical density (O.D.) value in $80{\mu}g/ml$. 2. The difference of ionic composition in DW, PBS and RPMI did not influence the adsorption amount of hFNIII 9-10.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.1
• /
• pp.61-64
• /
• 2000

A series of 5-oxaproline peptide derivatives was synthesized and evaluated for its ability to inhibit the prolyl 4-hydroxylase in vitro. Structure-activity studies show that the 5-oxaproline sequences, prepared by the 1,3-dipolar cycloaddition of the C-methoxycarbonyl-N-mannosyl nitrone in the presence of the ethylene, are more active than the corresponding proline derivatives. Prolyl 4-hydroxylase belongs to a family of $Fe^{2+}-dependent$ dioxygenase, which catalyzes the formation of 4-hydroxyproline in collagens by the hydroxylation of proline residues in -Gly-Xaa-Pro-Gly- of procollagen chains. In this paper we discover the more selective N-Cbz-Gly-Phe-Pro-Gly-OEt $(K_m\;=\;520\;{\mu}M)$ sequences which are showed stronger binding than others in vitro. Therefore, we set out to investigate constrained tetrapeptide that was designed to mimic the proline structure of pep tides for the development of prolyl 4-hydroxylase inhibitor. From this result, we found that the most potent inhibitor is N-Dansyl-Gly-Phe-5-oxaPro-Gly-OEt $(K_i\;=\;1.6\;{\mu}M)$. This has prompted attempts to develop drugs which inhibit collagen synthesis. Prolyl 4-hydroxylase would seem a particularly suitable target for antifibrotic therapy.

• Proceedings of the KOSOMBE Conference
• /
• v.1997 no.11
• /
• pp.333-336
• /
• 1997

The adhesion of activated and normal platelets to fibrinogen requires the receptor binding site of GPIIb/IIIa. These recognition sites exists in the A ${\alpha}$ chain(RGDS at 572-575 and RGDF at 95-98) and the carboxy-terminal of ${\gamma}$ chain (HHLGGAKQAGDV at 400-411) of fibrinogen. In this study, we developed RGDF-immobilized surface to detect the unctional state of platelet. RGDF-immobilized surface was prepared on the glass using photolithographic technology. Platelet adhesion to RGDF-immobilized surface was observed by staining platelets with mepacrine using a fluorescence microscope using mepacrine. Using the RGDF peptide of fragment E, we observed that the platelets pretreated with PGE1 interacted incompletely with RGDF-immobilized surface, whereas ADP activated platelets interacted with the surface extensively. These results show that the distinct selectivity of RGDF-immobilized micro-patterned surface can be used to detect the unctional state of platelets.

• Animal cells and systems
• /
• v.10 no.4
• /
• pp.197-201
• /
• 2006

Diaminopimelate decarboxylase (DAPDC, EC 4.1.1.20) catalyzes the conversion of diaminopimelate into lysine (Lys), which is the last step in Lys biosynthetic pathway. The genes for DAPDC have been reported in many bacteria, and more recently in Arabidopsis. Here we report characterization of a gene for DAPDC from rice (OsDAPDC). Sequence analysis of a cDNA clone revealed a full-length open reading frame for OsDAPDC that encoded 490 amino acids, approximately 53.2 kDa protein. The OsDAPDC protein contains a consensus binding site for pyridoxal-5'-phosphate as a cofactor and has a sequence at the amino terminus that resembles a transit peptide for localization to plastids, similar to that of Arabidopsis. Single gene encoding DAPDC was found in chromosome II in rice. The predicted amino acid sequence of OsDAPDC is highly homologous to that of the enzymes for DAPDC encoded by lysA of many bacteria. Expression of OsDAPDC in lysA mutants of Escherichia coli shows that the gene is able to functionally complement the mutants. These results suggest that OsDAPDC encodes a protein for diaminopimelate decarboxylase in rice.

• Animal cells and systems
• /
• v.16 no.2
• /
• pp.114-120
• /
• 2012

In echinoderms, the SALMFamide neuropeptides sharing the SxL/FxFamide motif seem widespread throughout the phylum and may be important signalling molecules that mediate various physiological functions. Recent identification of S1 and its analogues, MagS3 and MagS4, along with the S2 analogue, MagS2 from the starfish $Marthasterias$ $glacialis$, indicated that SALMFamides in the class Asteroidea are more diverse than previously thought. Further, isolation of the neuropeptides from the radial nerve cord and studies on pharmacological actions of the neuropeptides on the cardiac stomach warrant studies on the tissue distributions of these peptides in both the nervous and digestive systems. In the present study, antisera raised against an S1 analogue, KYSALMFamide, and an S2 analogue, KYSGLTFamide, were used to localize the distribution patterns of the S1- and S2-like immunoreactivities (S1-IR/S2-IR) in the nervous and digestive systems of the starfish. In the nervous system, cell bodies in the ectoneural part were immunostained for both S1 and S2 peptides, while in the digestive system, the basiepithelial plexus and mucosal cell bodies were immunoreactive. These immunocytochemical data support the notion that the SALMFamides may play a neuroendocrine role in mediating feeding behaviour of the starfish. Further studies including identification of peptide binding sites and differential expression pattern of mRNAs encoding the peptides are required to elucidate their physiological functions.

• Journal of Microbiology and Biotechnology
• /
• v.2 no.1
• /
• pp.56-65
• /
• 1992

The complete nucleotide sequence of the Bacillus circulans F-2 RSDA gene, coding for raw starch digesting a-amylase (RSDA), has been determined. The RSDA structure gene consists of an open reading frame of 2508 bp. Six bp upstream of the translational start codon of the RSDA is a typical gram-positive Shine-Dalgarno sequence and the RSDA encodes a preprotein of 836 amino acids with an Mr of 96, 727. The gene was expressed from its own regulatory region in E. coli and two putative consensus promoter sequences were identified upstream of a ribosome binding site and an ATG start codon. Confirmation of the nucleotide sequence was obtained and the signal peptide cleavage site was identified by comparing the predicted amino acid sequence with that derived by N-terminal analysis of the purified RSDA. The deduced N-terminal region of the RSDA conforms to the general pattern for the signal peptides of secreted prokaryotic proteins. The complete amino acid sequence was deduced and homology with other enzymes was compared. The results suggested that the Thr-Ser-rich hinge region and the non-catalytic domain are necessary for efficient adsorption onto raw substrates, and the catalytic domain (60 kDa) is necessary for the hydrolysis of substrates, as suggested in previous studies (8, 9).

• IMMUNE NETWORK
• /
• v.11 no.6
• /
• pp.399-405
• /
• 2011

Background: Endogenous uveitis is a chronic inflammatory eye disease of human, which frequently leads to blindness. Experimental autoimmune uveoretinitis (EAU) is an animal disease model of human endogenous uveitis and can be induced in susceptible animals by immunization with retinal antigens. EAU resembles the key immunological characteristics of human disease in that both are $CD4^+$ T-cell mediated diseases. Dendritic cells (DCs) are specialized antigen-presenting cells that are uniquely capable of activating naive T cells. Regulation of immune responses through modulation of DCs has thus been tried extensively. Recently our group reported that donor strain-derived immature DC pretreatment successfully controlled the adverse immune response during allogeneic transplantation. Methods: EAU was induced by immunization with human interphotoreceptor retinoid-binding protein (IRBP) $peptide_{1-20}$. Dendritic cells were differentiated from bone marrow in the presence of recombinant GM-CSF. Results: In this study, we used paraformaldehyde-fixed bone marrow-derived DCs to maintain them in an immature state. Pretreatment with fixed immature DCs, but not fixed mature DCs, ameliorated the disease progression of EAU by inhibiting uveitogenic $CD4^+$ T cell activation and differentiation. Conclusion: Application of iBMDC prepared according to the protocol of this study would provide an important treatment modality for the autoimmune diseases and transplantation rejection.

• Bulletin of the Korean Chemical Society
• /
• v.20 no.3
• /
• pp.301-306
• /
• 1999

Hepatitis C virus (HCV) has been known to be an enveloped virus with a positive strand RNA genome and the major agent of the vast majority of transfusion associated cases of hepatitis. For viral replication, HCV structural proteins are first processed by host cell signal peptidases and NS2/NS3 site of the nonstructural protein is cleaved by a zinc-dependent protease NS2 with N-terminal NS3. The four remaining junctions are cleaved by a separate NS3 protease. The solution conformations of NS4B/5A, NS5A/5B substrates and NS5A/5B inhibitor have been determined by two-dimensional nuclear magnetic resonance (NMR) spectroscopy. NMR data suggested that the both NS5A/5B substrate and inhibitor appeared to have a folded tum-like conformation not only between P1 and P6 position but also C-terminal region, whereas the NS4B/5A substrate exhibited mostly extended conformation. In addition, we have found that the conformation of the NS5A/5B inhibitor slightly differs from that of NS5A/5B substrate peptide, suggesting different binding mode for NS3 protease. These findings will be of importance for designing efficient inhibitor to suppress HCV processing.