• 생명과학회지
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• 제23권3호
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• pp.347-354
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• 2013

선천성 면역은 감염성 매개체를 자기(self)로부터 변별할 수 있다. 선천성 면역은 감염 초기에 숙주인 자기를 보호하는 인식분자와 효과인자들로 구성되어 있다. Mannose 결합 렉틴(Mannose-binding lectin, MBL)은 $Ca^{2+}$ 의존형 렉틴에 속하며, 콜라겐 유사 domain을 함유하는 C-type 렉틴으로 선천성 면역의 중요한 분자이다. 혈액내 낮은 MBL 농도는 면역결핍 증후군을 나타내며 감염에 대한 심각한 위험성을 초래한다. 사람의 MBL 결핍증은 coding 영역의 돌연변이에 의해 나타나며, 이 돌연변이의 영향을 연구하기 위해 쥐의 상동성 유전자인 MBL-A를 이용하고 있다. 돌연변이 MBL의 기능적, 세포 생리적 연구를 위해 선행연구에서 rat wild type MBL-A 유전자를 클로닝하였으며, 본 연구에서 이 유전자에 콜라겐 유사 domain에서 발견된 세 가지 돌연변이, R40C, G42D, G45E를 site-directed mutagenesis 방법으로 모두 도입하였다. 세 가지 돌연변이가 존재하는 MBL 단백질은 정상 MBL과 마찬가지로 세포 내에서 정상적으로 발현되었으며, 여전히 렉틴 기능을 가지고 있었다. 이는 세 가지 돌연변이가 렉틴 기능을 나타내는 C-말단 쪽의 carbohydrate recognition domain에는 구조적으로, 또한 기능적으로도 영향을 미치지 않는다는 결과이다. 그러나 이 돌연변이는 MBL 단백질이 세포 밖으로 분비되는 것을 방해하였으며, 그 결과로 소포체 내에 잔류하여 소포체 망상구조(endoplasmic reticulumn network)에 커다란 손상을 주며 비정상적인 형체를 초래하였다. 이 같은 결과는 돌연변이 MBL에 의해 나타난 세포 내 병리현상의 새로운 발견으로 향후 MBL의 구조 형성과 분비 연구에 기여를 할 것으로 생각된다.

• Journal of the korean society of oceanography
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• 제35권3호
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• pp.153-157
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• 2000

Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.35-39
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• 2001

A simple method for studying the lectin-like interactions between Helicobacter pylori and cultured human epithelial cell lines was developed using an enzyme-linked, biotin-streptavidin bacterial-adhesion assay. The present study suggests that this method is suitable for evaluating the participation of lectin interactions in the adhesion of H. pylori to cultured HeLa S3 and Kato III cells, both fixed and glycosidase-treated cells, as well as assessing glycoconjugated binding inhibition studies. The time-course and dose-dependent kinetics of the biotin-labeled H. pylori adhesion th the formaldehyde-fixed Hela S3 and Kato III cell lines exhibited saturation. In addition, the binding of the biotin-labeled H. pylori to the formaldehyde-fixed cultured cells was partially blocked by pre-incubation with glycoconjugates and polyclonal antibodies against a heparan sulfate binding protein from H. pylori.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.334-340
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• 2005

Lectin is a cell-agglutinating and carbohydrate-binding protein present in many plants. The lectin of Canavalia ensiformis shoot with specific affinity for D-glucose was purified by affinity chromatography using Sephadex G-100, and some of its biochemical characterizations were studied. Lectin was purified 8.87-fold and exhibited final specific activity of 225.74 units/mg protein with a $2.3\%$ yield. SDS-PAGE analysis demonstrated that the purified shoot lectin exists as a tetramer of 102 kD, composed of two subunits with molecular weight of 29 and 22 kD. The purified lectin was observed to agglutinate rabbit blood cell. The optimal temperature for the activity of this lectin was $40^{\circ}C$, and this lectin was relatively stable to heat with the highest activity at $50{\~}60^{\circ}C$. The maximal activity was observed at pH 7.2.

• 약학회지
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• 제50권3호
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• pp.166-171
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• 2006

A calcium-dependent sialic acid-binding lectin has been isolated by thyroglobulin-affinity chromatography from the coastal crab Macrophthalmus Japonicus. This lectin, Macrophthalmus Japonicus lectin (MJL), was eluted with 50mM Tris-HCl, 0.3 M NaCl, 10 mM EDTA, and the recovery yield from the crude protein extract was about 5.6%. The molecular weight of MJL was estimated as 65 kDa in SDS-PAGE both under reducing and non-reducing conditions. MJL induced an agglutination reaction in rabbit, rat, and mouse erythrocytes, but not in human ABO types. This activity was effectively inhibited by sialoglycoproteins such as fetuin, bovine submaxillary mucin, and thyroglobulin.

• BMB Reports
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• 제40권3호
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• pp.358-367
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• 2007

A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95 $\mu$g/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7 $\pm$ 6.8), Bre-04 (41.5 $\pm$ 4.8), and Lu-04 (11.4 $\pm$ 0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with $TC_50$ and $EC_50$ of 5.176 mg/ml and 3.054 $\mu$g/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.

• Journal of Microbiology and Biotechnology
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• 제33권5호
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• pp.698-705
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• 2023

Rapid diagnosis of methicillin-resistant Staphylococcus aureus (MRSA) is essential for guiding clinical treatment and preventing the spread of MRSA infections. Herein, we present a simple and rapid MRSA screening test based on the aggregation effect of mannose-binding lectin (MBL)-conjugated gold nanoparticles (AuNP), called the MRSA probe. Recombinant MBL protein is a member of the lectin family and part of the innate immune system. It can recognize wall teichoic acid (WTA) on the membrane of MRSA more specifically than that of methicillin-sensitive Staphylococcus aureus (MSSA) under optimized salt conditions. Thus, the MRSA probe can selectively bind to MRSA, and the aggregation of the probes on the surface of the target bacteria can be detected and analyzed by the naked eye within 5 min. To demonstrate the suitability of the method for real-world application, we tested 40 clinical S. aureus isolates (including 20 MRSA specimens) and recorded a sensitivity of 100%. In conclusion, the MRSA probe-based screening test with its excellent sensitivity has the potential for successful application in the microbiology laboratory.

• 한국식품영양과학회지
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• 제23권3호
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• pp.515-519
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• 1994

지금까지 발표된 본 연구실의 논문 중 팥콩 lectin(KBL)과 토란 lectin(TTL)에 관하여, 물리화학적인 특성과 독활성을 중심으로, 각 논문의 자료를 토대로 비료 정리하였다.

• 생명과학회지
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• 제18권11호
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• pp.1543-1550
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• 2008

실내 환경 오염원으로 각종 암 발병과 관련이 있는 흡연이 피부 피지샘의 glycoconjugates에 미치는 영향을 lectin 조직화학으로 연구하였다. Sprague-Dawley계 수컷 흰쥐에 1일 1회 10분간 1일군, 2일군, 3일군 및 5일군으로 나누어 직?간접 흡연 방법으로 폭로시켜 피지샘 형태는 PAS 염색, 피지샘의 glycoconjugates 양상을 9종의 biotinylated lectin (DBA, SBA, PNA, BSL-1, WGA, RCA-1, UEA-1, Con A 및 LCA)으로 연구하였다. 대조군에 비해 흡연군에서는 피지샘꽈리의 크기 감소, 피지샘꽈리 상부의 파괴, 중앙부 피지세포의 공포화 및 미성숙 피지샘꽈리가 관찰되었다. 대조군에서 피지샘 glycoconjugates에는 염색성에 차이가 있으나 기저세포는 BSL-1, PNA 및 WGA에만 반응하고 BSL-1의 반응이 더 강하며, 피지세포는 PNA, WGA, Con A, BSL-1 및 SBA에만 반응하고 PNA 및 Con A의 반응이 더 강했다. 흡연군의 렉틴 반응에서 특이적인 변화가 관찰되었다. 흡연군의 피지샘 기저세포의 PNA 반응은 증가되었고, BSL-1는 감소되었다가 회복되었으며, WGA는 소실되었다. 피지세포의 Con A는 증가하였으나 PNA, BSL-1, WGA 및 SBA는 감소 또는 소실되었다. 대조군에 관찰되지 않던 흡연군의 미성숙 샘꽈리 PNA, on A 및 BSL-1이 반응하였으며 PNA 및 Con A의 반응이 더 현저하였다. 결론적으로 흡연은 피지샘의 형태 및 glycoconjugates 대사에 심한 영향을 미치는 것으로 사료된다.

• 분석과학
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• 제13권5호
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• pp.624-629
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• 2000

Mistletoe lectin I(ML I)에 대한 간단하고 빠른 용액상 효소결합 분석법을 렉틴의 당 특이성을 이용하여 개발하였다. ML I에 특이성을 가지고 있는 D-galactose를 사용하였으며, 용액상 분석법의 효소로는 malate dehydrogenase(MDH)를 사용하였다. 분석신호물질로 사용되는 MDH-galactose 접합체는 isothiocyanate 방법을 통해 합성하였으며, 이 접합제는 thiourea 결합을 하고 있다. ML I의 존재하에, ML I은 D-galactole와의 특이 인식결합을 통해 MDH-galartose 접합체의 활동도를 억제한다. 그러므로, 존재하는 ML I의 농도는 MDH-galactose 접합제의 촉매활동도의 억제도에 비례하게 된다. 따라서, 본 용액상 효소결합 분석법을 통하여 ${\mu}g/mL$ 수준의 ML I의 측정이 분석 시간 10분 이내에 가능하였다.