• Journal of Microbiology and Biotechnology
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• 제25권1호
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• pp.119-126
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• 2015

Among the various human growth factors, epidermal growth factor (hEGF, consisting of 53 amino acids) has various effects on cell regeneration, stimulation of proliferation, migration of keratinocytes, formation of granulation tissues, and stimulation of fibroblast motility, which are important for wound healing. Owing to their multiple activities, EGFs are used as pharmaceutical and cosmetic agents. However, their low productivity, limited target specificity, and short half-life inhibit their application as therapeutic agents. To overcome these obstacles, we fused the collagen-binding domain (CBD) of Vibrio mimicus metalloprotease to EGF protein. About 18 or 12 amino acids (aa) (of the 33 total amino acids), which were essential for collagen-binding activity, were combined with the N- and C-termini of EGF. We constructed, expressed, and purified EGF (53 aa)-CBD (18 aa), EGF (53 aa)-CBD (12 aa), CBD (18 aa)-EGF (53 aa), and CBD (12 aa)-EGF (53 aa). These purified recombinant proteins increased the numbers of cells in treated specimens compared with non-treated specimens and control hEGF samples. The collagen-binding activities were also evaluated. Furthermore, CBD-hybridized hEGF induced phosphorylation of the EGF receptor. These results suggested that these fusion proteins could be applicable as small therapeutic agents in wound tissue healing.

• BMB Reports
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• 제52권10호
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• pp.601-606
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• 2019

Arginine methylation plays crucial roles in many cellular functions including signal transduction, RNA transcription, and regulation of gene expression. Protein arginine methyltransferase 8 (PRMT8), a unique brain-specific protein, is localized to the plasma membrane. However, the detailed molecular mechanisms underlying PRMT8 plasma membrane targeting remain unclear. Here, we demonstrate that the N-terminal 20 amino acids of PRMT8 are sufficient for plasma membrane localization and that oligomerization enhances membrane localization. The basic amino acids, combined with myristoylation within the N-terminal 20 amino acids of PRMT8, are critical for plasma membrane targeting. We also found that substituting Gly-2 with Ala [PRMT8(G2A)] or Cys-9 with Ser [PRMT8(C9S)] induces the formation of punctate structures in the cytosol or patch-like plasma membrane localization, respectively. Impairment of PRMT8 oligomerization/dimerization by C-terminal deletion induces PRMT8 mis-localization to the mitochondria, prevents the formation of punctate structures by PRMT8(G2A), and inhibits PRMT8(C9S) patch-like plasma membrane localization. Overall, these results suggest that oligomerization/dimerization plays several roles in inducing the efficient and specific plasma membrane localization of PRMT8.

• Nutrition Research and Practice
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• 제12권3호
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• pp.208-214
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• 2018

BACKGROUND/OBJECTIVES: The aim of this study was to evaluate the biological and sleep-promoting effects of combined ${\gamma}$-aminobutyric acid (GABA) and 5-hydroxytryptophan (5-HTP) using caffeine-induced sleepless fruit flies, ICR mice, and Sprague-Dawley rats. MATERIALS/METHODS: Video-tracking analysis was applied to investigate behavioral changes of Drosophila melanogaster. Pentobarbital-induced sleep test and electroencephalogram (EEG) patterns were used for analysis of sleep latency, duration, and quantity and quality of sleep in vertebrate models. RESULTS: Administration of combined GABA/5-HTP could significantly reverse the caffeine induced total distance of flies (P < 0.001). Also, individually administered and combined GABA/5-HTP significantly increased the total sleeping time in the caffeine-induced sleepless ICR mice (P < 0.001). In the caffeine-induced sleepless SD-rats, combined GABA/5-HTP showed significant differences in sleep quality between individual amino acid administrations (P < 0.05). CONCLUSIONS: Taken together, we identified inhibitory effects of combined GABA/5-HTP in locomotor activity, sleep quantity and quality in caffeine-induced sleepless models, indicating that combined GABA/5-HTP may be effective in patients with insomnia by providing sufficient sleep.

• Applied Biological Chemistry
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• 제20권1호
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• pp.130-135
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• 1977

가스 크로마토와 가스크로마-토질량분석기에 의하여 수종의 수산기 함유 아미노산의 치환에 스터화반응을 고찰하고 이들의 fragmentation기작을 설명 했다.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.239-249
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• 1999

The role of amino acids in culture media for IVF-ET was examined in a total of 76 cycles. Patients received clomiphene citrate (CC) followed by hMG or GnRH-a combined with gonadotropins (FSH/hMG) for controlled ovarian hyperstimulation. Severe male (<$4{\times}10^6$ motile sperm) or age factor (>39 y) patients were excluded in this study. Pregnancy was classified as clinical if a gestational sac or fetal cardiac activity was seen on ultrasound. No significant differences were found in age, duration of infertility, follicle size, the level of $E_2$ on the day of hCG injection, the mean number of oocytes retrieved, total motile sperm count, fertilization rate and the mean number of embryos transferred between bHTF (without amino acids) and mHTF (with amino acids) groups. However, total ampules of gonadotropins were higher (p<0.01) in mHTF group than bHTF group. Significantly (p<0.05) more clinical pregnancies were recorded in mHTF group (13/30) compared with bHTF group (9/46). The multiple pregnancy rates were 11.1% in bHTF group and 7.7% in mHTF group. There were one ectopic pregnancy in mHTF group and one heterotopic pregnancy in bHTF group. Abortion rates were 22.2% in bHTF group and 7.7% in mHTF, respectively. The ongoing pregnancy or livebirth rate was significantly (p<0.05) higher in mHTF group (12/30) than bHTF group (7/46). These results suggest that the addition of amino acids in culture media is essential for culture of zygotes in vitro and adjustment of energy substrates in phosphate-free culture media appears to be beneficial for human IVF-ET procedure.

• Asian-Australasian Journal of Animal Sciences
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• 제28권4호
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• pp.485-493
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• 2015

This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen microorganisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance.

• 한국식품과학회지
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• 제3권1호
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• pp.57-63
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• 1971

1. 5섯가지 젖산균을 대두유에 접종배양 하여 산 생성도를 비교 하였드니 Str. thermophilus 와 Str. diacetilactis가 Str. lactis, Str. cremoris 및 L. bulgaricus보다 높았다. 2. 대두유와 탈지 분유를 7 : 3의 비로 혼합하여 젖산균 발효를 하였을 때에 침전물의 수획량이 가장 많고 또한 그의 단백질 함량이 가장 많으며 수분의 량이 적고 견고한 curd를 만들수 있음을 알았다. 3. 대두유와 탈지분유를 7 : 3의 비로 혼합한 것에 젖산균 starter로 Str. thermophilus를 첨가하고 Pen. caseicolum을 접종하여 제조한 치-즈를 3주간 숙성$(15^{\circ}C,\;RH:\;85{\sim}90%)$ 하였드니 숙성 7일째부터 Pen. caseicolum의 생육이 왕성하게 되며 pH가 상승함과 동시에 proteolytic activity는 14일째에 최고치에 도달하였다, 또 total water soluble nitrogen, water soluble protein nitrogen 및 amino-N도 숙성 7일째부터 증가하기 시작하여 숙성 21일째는 이들의 값이 각각 total nitrogen의 52%, 32% 및 14%에 이르렀다. 그리고 이차원 paper chromatography에 의하여 3주간 숙성한 치-즈에서 17종의 아미노산을 검출 하였다. 또 3주간 숙성한 후 제품의 일반성분 조성은 수분 63.2%, 조단백질 17.5%, 조지방 13.2%, 조회분 2.8% 및 식염 2.5%였다.

• 한국식품과학회지
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• 제54권1호
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• pp.1-7
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• 2022

본 연구에서는 형광등 빛 조사 하에 각종 감광제와 아미노산의 조합에 의한 NBT의 환원 특성을 조사하였다. 기존 SOD 활성 측정에 이용된 Rb-Met 반응계는 증류수나 Tris보다는 PBS 용매 상에서 가장 우수한 효과를 나타냈다. 빛 조사 하에 Rb에 의한 NBT 환원을 위해서는 Met과 His 등의 아미노산이 필요하며, 감광제로서는 isoalloxazine 계열의 Rb와 thiazine 계열의 MB가 효과적인 환원 반응을 유도했다. 하지만 각종 감광제나 아미노산의 조합, 그리고 반응 용매에 따라 NBT 환원정도가 상이하며, 특히 Rb-Met in PBS와 MB-His in Tris 반응계가 가장 큰 반응을 유도하였다. Rb에 의해 유도된 NBT 환원반응은 SOD 및 gallic acid에 의해 효과적으로 저해되었으나, Tris 상에서 MB-His에 의한 NBT 환원은 SOD 및 gallic acid에 의한 저해효과가 미미하여 Rb와는 다른 기작에 의해 NBT 환원이 유도되는 것으로 보인다. 본 결과는 감광제와 아미노산에 따라 빛 조사 하에 다양한 상호작용이 발생하며, NBT 환원을 유도하는데 superoxide anion 뿐만 아니라 다른 요인도 관여할 수 있음을 시사한다.

• The Korean Journal of Physiology and Pharmacology
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• 제3권1호
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• pp.35-45
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• 1999

Excitatory amino acid (EAA) and substance P (SP) have been known to be primary candidates for nociceptive neurotransmitter in the spinal cord, and calcium ions are implicated in processing of the sensory informations mediated by EAA and SP in the spinal cord. In this study, we examined how $Ca^{2+}$ modified the responses of dorsal horn neurons to single or combined iontophoretical application of EAA and SP in the rat. All the LT cells tested responded to kainate, whereas about 55% of low threshold (LT) cells responded to iontophoretically applied NMDA. NMDA and kainate excited almost all wide dynamic range (WDR) cells. These NMDA- and kainate-induced WDR cell responses were augmented by iontophoretically applied EGTA, but suppressed by $Ca^{2+},\;Mn^{2+},$ verapamil and ${\omega}-conotoxin$ EVTA, effect of verapamil being more prominent and well sustained. $Ca^{2+}$ and $Mn^{2+}$ antagonized the augmenting effect of EGTA. On the other hand, prolonged spinal application of EGTA suppressed the response of WDR cell to NMDA. SP had triple effects on the spontaneous activity as well as NMDA-induced responses of WDR cells: excitation, inhibition and no change. EGTA augmented, but $Ca^{2+},\;Mn^{2+}$ and verapamil suppressed the increase in the NMDA-induced responses and spontaneous activities of WDR cells following iontophoretical application of SP. These results suggest that in the spinal cord, sensory informations mediated by single or combined action of EAA and SP can be modified by the change in calcium ion concentration.

• 한국축산식품학회지
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• 제33권4호
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• pp.474-480
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• 2013

This study was performed to investigate the effects of high pressure/high temperature (HPHT) treatment on the recovery efficiency and characteristics of porcine placenta hydrolysates. The placenta hydrolysates were characterized by solubility, free amino acid contents, gel electrophoresis, gel permeation chromatography (GPC) and amino acid composition. Placenta was treated at 37.5 MPa of pressure combined with various temperatures (150, 170, and $200^{\circ}C$) or various holding times (0, 30, and 60 min at $170^{\circ}C$). Insoluble raw placenta collagen was partially solubilized (> 60% solubility) by the HPHT treatment. Free amino group content of placenta collagen was increased from 0.1 mM/g collagen to > 0.3 mM/g collagen after HPHT treatment, reflecting partial hydrolysis of collagen. The molecular weight ($M_w$) distribution showed evidence of collagen hydrolysis by shifting of $M_w$ peaks toward low molecular weight when treated temperature or holding time was increased. Alanine (Ala), glycine (Gly), hydroxyproline (Hyp), and proline (Pro) contents increased after the HPHT treatments compared to a decrease in the others. In particular, the increase in Gly was obvious, followed by Hyp and Pro, reflecting that placenta hydrolysates were mainly composed of these amino acids. However, increasing temperature or holding time hardly affected the amino acid compositions. These results indicate that the HPHT treatment is advantageous to hydrolyze collagen derived from animal by-products.