• Restorative Dentistry and Endodontics
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• v.32 no.5
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• pp.459-468
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• 2007

Protein microarray or protein chips is potentially powerful tools for analysis of protein-protein interactions. APin cDNA was previously identified and cloned from a rat odontoblast cDNA library. The purpose of this study was to investigate the APin-protein interactions during ameloblast differentiation. Protein microarray was carried with recombinant APin protein and MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein were selected among 74 interacting proteins. Immortalized ameloblast cells (ALCs) were transfected with pCMV-APin construct and U6-APin siRNA construct. After transfection, the expression of the mRNAs for four proteins selected by protein micoarrays were assessed by RT-PCR. The results were as follows: 1. APin expression was increased and decreased markedly after its over-expression and inactivation, respectively. 2. Over-expression of the APin in the ALCs markedly down-regulated the expression of MEF2 and Aurora kinase A, whereas their expression remained unchanged by its inactivation. 3. Expression of BMPR-IB and EF-hand calcium binding protein were markedly increased by the over-expression of the APin in the ALCs, whereas expression of BMPR-IB remained unchanged and expression of EF-hand calcium binding protein was markedly decreased by its inactivation. These results suggest that APin plays an important role in ameloblast differentiation and mineralization by regulating the expression of MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein.

• Journal of Applied Biological Chemistry
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• v.53 no.3
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• pp.174-178
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• 2010

Calcium and iron binding peptides were prepared by enzymatic hydrolysis and ultrafiltration of rice bran protein (RBP), which was isolated from defatted rice bran by phytase and xylanase treatment and ultrasonication. The isolated RBP had a molecular weight in the range of 10-66 kDa. The extracted proteins were hydrolyzed using Flavourzyme for 6 hr. After ultrafiltration under 5 kDa as molecular weight, the peptides were fractionated into 4 peaks by Sephadex G-15 gel permeation chromatography, and each fraction was determined for calcium and iron binding activity. As the result, Fl and F2 fractions were the best candidate for calcium and iron chelation, respectively. These results suggest that the calcium and iron binding peptides can be used as functional food additives in food industry.

• Molecules and Cells
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• v.41 no.2
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• pp.103-109
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• 2018

Calcium ions are involved in the regulation of diverse cellular processes. Fourteen genes encoding calcium binding proteins have been identified in Dictyostelium. CBP7, one of the 14 CBPs, is composed of 169 amino acids and contains four EF-hand motifs. Here, we investigated the roles of CBP7 in the development and cell migration of Dictyostelium cells and found that high levels of CBP7 exerted a negative effect on cells aggregation during development, possibly by inhibiting chemoattractant-directed cell migration. While cells lacking CBP7 exhibited normal development and chemotaxis similar that of wild-type cells, CBP7 overexpressing cells completely lost their chemotactic abilities to move toward increasing cAMP concentrations. This resulted in inhibition of cellular aggregation, a process required for forming multicellular organisms during development. Low levels of cytosolic free calcium were observed in CBP7 overexpressing cells, which was likely the underlying cause of their lack of chemotaxis. Our results demonstrate that CBP7 plays an important role in cell spreading and cell-substrate adhesion. cbp7 null cells showed decreased cell size and cell-substrate adhesion. The present study contributes to further understanding the role of calcium signaling in regulation of cell migration and development.

• Molecules and Cells
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• v.19 no.3
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• pp.382-390
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• 2005

Calcium-binding proteins are thought to play important roles in calcium buffering. The present study investigated the effects of ischemia and reperfusion on calbindin D28K, calretinin, and parvalbumin immunoreactivity in the ganglion cell layer of the rabbit. Rabbits were administered ischemic damage by increasing the intraocular pressure. After 60 and 90 min of ischemia, reperfusion (7 d) was allowed to occur. The b-wave of the electroretinogram (ERG) was reduced by more than 50% and almost 80% in retina given ischemia for 60 and 90 min, respectively. The oscillatory potential (OPs) wave was reduced approximately 50% at 60 min ischemia and 70% at 90 min ischemia. In both normal and ischemic-treated retina, calcium-binding protein immunoreactivity was seen in many cells in the ganglion cell layer. In eyes subjected to 60 min ischemia, there was a decrease of the density of calbindin D28K- (8.29%), calretinin- (14.44%), and parvalbumin- (26.83%) immunoreactive (IR) cells compared to the control retina. In eyes subjected to 90 min ischemia, there was a higher decrease of the density of calbindin D28K- (18.48%), calretinin- (33.59%), and parvalbumin- (54.26%) IR cells than at 60 min. Some calcium-binding protein-IR neurons, especially calretinin-IR neurons, showed aggregations that were abnormally packed together in retina subjected to ischemia for 90 min. The results show that calbindin D28K-, calretinin-, and parvalbumin-IR cells in the ganglion cell layer are susceptible to ischemic damage and reperfusion. The degree of reduction varied among different calcium-binding proteins and ischemic damage times. These results suggest that calbindin D28K-containing neurons are less susceptible to ischemic damage than calretinin- and parvalbumin-containing neurons in the ganglion cell layer of rabbit retina.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.139-146
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• 2001

This study was carry out to identify the distribution of calcium binding proteins; calbindin(CB), calretinin(CR) and parvalbumin(PA) in the suprachiasmatic nucleus(SCN) of the Korean native goat by immunohistochemical methods. The expression of substance P(SP), calcitonin gene-related peptide(CG-RP), neuropeptide Y(NPY), vasoactive intestinal polypeptide(VIP) and galanin(GAL) were also investigated. CR-immunoreactivity was found in both of the cell bodies and fibers in the SCN, which the CB-immunoreactivity was observed only in the fibers. The immunoreactivity for VIP was observed in both the cell bodies and fibers, but SP-, NPY, GAL-immunoreactivities were only found in the fibers. CGRP-immunoreactivity was not seen in cell body and fibers. These results suggest that VIP, SP, NPY and GAL play a neuromodulatory or/ and neurotransmitter roles in cooperation with CB and CR in SCN of the Korean native goat.

• Biomolecules & Therapeutics
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• v.17 no.2
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• pp.125-132
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• 2009

Calbindin-$D_{9k}$ (CaBP-9k), a cytosolic calcium-binding protein, is expressed in a variety of tissues, i.e., the duodenum, uterus, placenta, kidney and pituitary gland. Duodenal CaBP-9k is involved in intestinal calcium absorption, and is regulated at transcriptional and post-transcriptional levels by 1,25-dihydroxyvitamin D3, the hormonal form of vitamin D, and glucocorticoids (GCs). Uterine CaBP-9k has been implicated in the regulation of myometrial action(s) through modulation of intracellular calcium, and steroid hormones appear to be the main regulators in its uterine and placental regulation. Because phenotypes of CaBP-9k-null mice appear to be normal, other calcium-transporter genes may compensate for its gene deletion and physiological function in knockout mice. Previous studies indicate that CaBP-9k may be controlled in a tissue-specific fashion. In this review, we summarize the current information on calcium homeostasis related to CaBP-9k gene regulation by GCs, vitamin D and its receptors, and its molecular regulatory mechanism. In addition, we present related data from our current research.

• BMB Reports
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• v.40 no.3
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• pp.302-314
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• 2007

N type calcium channels (CaV2.2) play a key role in the gating of transmitter release at presynaptic nerve terminals. These channels are generally regarded as parts of a multimolecular complex that can modulate their open probability and ensure their location near the vesicle docking and fusion sites. However, the proteins that comprise this component remain poorly characterized. We have carried out the first open screen of presynaptic CaV2.2 complex members by an antibody-mediated capture of the channel from purified rat brain synaptosome lysate followed by mass spectroscopy. 589 unique peptides resulted in a high confidence match of 104 total proteins and 40 synaptosome proteome proteins. This screen identified several known CaV2.2 interacting proteins including syntaxin 1, VAMP, protein phosphatase 2A, $G_{o\alpha}$, G$\beta$ and spectrin and also a number of novel proteins, including clathrin, adaptin, dynamin, dynein, NSF and actin. The unexpected proteins were classified within a number of functional classes that include exocytosis, endocytosis, cytoplasmic matrix, modulators, chaperones, and cell-signaling molecules and this list was contrasted to previous reports that catalogue the synaptosome proteome. The failure to detect any postsynaptic density proteins suggests that the channel itself does not exhibit stable trans-synaptic attachments. Our results suggest that the channel is anchored to a cytoplasmic matrix related to the previously described particle web.

• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.17-17
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• 1996

Annexin I is a member of the annexin family of calcium dependent phospholipid binding proteins and has anti-inflammatory activity by inhibiting phospholipase A$_2$ (PLA$_2$). Recent X-ray crystallographic study of annexin I identified six Ca$\^$2+/ binding sites, which was different types (type II, III) from the well-known EF-hand motif (type I). (omitted)

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.435-447
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• 1999

The localization of substance P(SP), calcitonin gene-related peptide(CGRP) and three calcium binding proteins, calbindin D-28k(CB), calretinin(CR) and parvalbumin(PA) was immunohistochemically examined in the myenteric and submucous plexuses of Korean native goat duodenum. In the neurons of myenteric and submucous plexuses of duodenum, immunoreactivities of SP, CGRP and CB were confirmed in both nerve cell bodies and fibers. In contrast, CR immunoreactivity was found only in nerve fibers of myenteric plexuses, while PA immunoreactivity was found only in nerve cell bodies of submucous plexuses. In the inner circular muscle layer, dense SP-like immunoreactive fibers were prominent but only a few CGRP-like immunoreactivities were observed. Most of SP- and CGRP-like immunoreactive neurons of both plexuses colocalized with CB. This result showed that SP and CGRP may have a important role for the movement of small intestine. The colocalizations of CB with SP or CGRP in myenteric and submucous plexuses suggest that CB may serve neuromodulatory role for SP- and CGRP-immunoreacted neurons on the movement of intestinal wall.

• Current Research on Agriculture and Life Sciences
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• v.32 no.1
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• pp.34-42
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• 2014

Soy and fermented soy are popular and recognized as a health food among Koreans. Since soy proteins are known to be protease resistant, even to pepsin and pancreatin, it is hypothesized that soy proteins may interact with the intestinal tract and trigger certain physiological reactions. To test this hypothesis, mice were fed diets supplemented with soy, Chunkukjang, or casein. The differentially expressed proteins were analyzed using 2-D gels and identified by peptide mass fingerprinting using mass spectrometry. The majority of the differentially expressed proteins could be functionally grouped into metabolic enzymes and calcium-binding proteins. The differential protein expression by the soy-fed groups was also verified based on a representative protein, tropomyosin, using a Western blotting analysis. In addition, the soy-fed groups exhibited a taller villi structure. Therefore, this study suggests that soy proteins can be an effective nutrient and physiological stimulant for the intestines.