• KSBB Journal
• /
• 제23권1호
• /
• pp.48-53
• /
• 2008

토마토의 locular fluid로부터 최종적으로 Sephadex G-200 affinity chromatography에 의해 lectin을 분리한 다음, 이들의 분자량, 적혈구 응집력, 혈액특이성, 열 안정성, 최적 온도 및 pH 안정성의 생화학적 성질을 연구하였다. SBS-PAGE의 결과, 분자량이 39 kDa와 23 kDa로서 각각 2개의 subunit로 구성된 124 kDa의 분자량을 가지는 tetramer이다. 트립신으로 처리된 사람의 A, B, O, AB형의 혈액을 사용하여 각각의 혈구응집반응을 확인한 결과, A, B, O, AB형 모두에서 응집반응이 일어났으며, 이 중 B형 혈액에서 가장 높은 활성을 나타냈으며, A와 O형은 중간, AB형은 가장 낮은 활성을 보였다. 분리된 토마토 locular fluid의 최적반응 온도는 $50^{\circ}C$로서, 가장 높은 $70^{\circ}C$를 포함하는 $40-80^{\circ}C$에서 열 안정성을 보였으며, 이의 최적 pH는 7.0이다.

• 한국어병학회지
• /
• 제25권1호
• /
• 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.

• 생명과학회지
• /
• 제17권2호통권82호
• /
• pp.254-259
• /
• 2007

Neutral saline 추출, ammonium sulfate 침전 및 Sephadex G-200을 사용한 affinity chromatography 과정을 통해 방울토마토 열매에서 분리된 lectin의 생화학적 특성을 연구하였다. 트립신을 처리한 사람의 ABO형 적혈구 모두에서 응집반응이 일어났으며, 이 중 B형 적혈구에서 가장 높은 응집반응이 관찰되었다. 전기영동 분석에 의해 분자량 10.7 kDa의 강도가 높은 밴드가 확인되었다. 분리된 lectin의 최적반응온도는 $40^{\circ}C$이며, 40-60$^{\circ}C$ 범위에서 열에 대해 안정하였다. 또한 최적 pH는 7.2로 조사되었다.

• 생명과학회지
• /
• 제23권3호
• /
• pp.347-354
• /
• 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.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제6권1호
• /
• pp.99-102
• /
• 2003

A cDNA of novel immulectin homologue (BmIML), a C-type lectin, was cloned from the silkworm, Bombyx mori. The immulectin cDNA is an open reading frame of 921 bp encoding 307 amino acid residues. The deduced amino acid sequence from the BmIML cDNA contains two C-type carbohydrate recognition domains (CRDs). The BmIML was most similar (61 % protein sequence identity) to the M. sexta immulectin-1, whereas BmIML showed relatively lower identity to the B. mori lipopolysaccharide-binding protein (25% protein sequence identity). These features of BmIML indicate that BmIML is a novel member of C-type lectin superfamily. Northern blot analysis revealed that the BmIML is specifically expressed in the fat body of B. moli larvae.

• 한국어업기술학회:학술대회논문집
• /
• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
• /
• pp.530-530
• /
• 2000

The host defense system or immune system of all modern animals has their roots in very ancient organisms. After analyzing literature data concerning properties of invertebrates and vertebrates lectins we suggest that mechanism of mannans recognition may exist in marine invertebrates, as a universal mechanism for homeostasis maintenance and host defense, and mannan-binding lectins family of vertebrates has ancient precursor, as was shown for another S-type lectins family. We carried out the screening of mannan-binding type lectin among different species of echinoderms inhabiting in Piter the Grate Bay, the sea of Japan. As a result, the C-type lectins (SJL-32) specific for high mannose glycans was isolated from the coelomic plasma of the sea cucumbers Stichopus japonicus by ion-exchange chromatography on a DEAE-Toyopearl 650M, affinity chromatography on a mannan-Sepharose 6B and gel filtration on a Sephacryl S-200. SJL-32 is homodimer with molecular mass about 32 kDa on SDS-PAGE under non-reducing conditions. Protein part of the lectin has high conteins Asn, Glu, Ser. Hemagglutination of trypsin-treated O blood group human erythrocytes by SJL-32 was competitively inhibited by high-branched -D-mannan composed of -1,2 and -1,6 linked D-mannopyranose residues. In contrast, a variety of mono-, oligo-, and polysaccharides composed of residues of galactose and fucose showed absence or little inhibitory activities. The lectin activity strong depends on Ca2+ concentration, temperature and pH. Monospecific polyclonal antibodies were obtained to the lectin. As was shown by ELISA assay, antibodies to SJL-32 cross-reacted with human serum mannan-binding lectin. This data allows making conclusion about common antigenic determinants and structural homology of both lectins. In our opinion, SJL-32 belongs to evolutionary high conservative mannan-binding lectins (MBLs) family and takes part in the host defense against pathogenic microorganisms.

• 한국식품과학회지
• /
• 제43권1호
• /
• pp.72-76
• /
• 2011

The adjuvant effects of Korean mistletoe lectin-C (KML-C) were investigated following the oral administration of KML-C with ovalbumin (OVA) as an antigen. Mice were orally immunized with OVA alone or admixed with various doses of KML-C or cholera toxin (CT), and the titer of OVA-specific antibody in the serum and mucosal secretions were determined. OVA+KML-C-treated mice showed high titers of IgA specific to CT in mucosal secretions. The antibody titers in the serum of OVA+KML-C-treated mice were comparable to those in the serum of OVA+CT-treated mice. When mice were immunized with OVA+KML-C or with CT alone and subsequently injected with OVA on the footpads after the primary immunization, they showed a more significant increase in delayed-type hypersensitivity reactions than when they were administered CT alone. These results suggest that KML-C is a potent immunoadjuvant that enhances both humoral and cellular immunity by the mucosal immune system.

• Parasites, Hosts and Diseases
• /
• 제45권4호
• /
• pp.255-266
• /
• 2007

The protective effect of the Synadenium carinatum latex lectin (ScLL), and the possibility of using it as an adjuvant in murine model of vaccination against American cutaneous leishmaniasis, were evaluated. BALB/c mice were immunized with the lectin ScLL (10, 50, 100$[\mu}g$/animal) separately or in association with the soluble Leishmania amazonensis antigen (SLA). After a challenge infection with $10^6$ promastigotes, the injury progression was monitored weekly by measuring the footpad swelling for 10 weeks. ScLL appeared to be capable of conferring partial protection to the animals, being most evident when ScLL was used in concentrations of 50 and 100${\mu}g$/animal. Also the parasite load in the interior of macrophages showed significant reduction (61.7%) when compared to the control group. With regard to the cellular response, ScLL 50 and 100 ${\mu}g$/animal stimulated the delayed-type hypersensitivity (DTH) reaction significantly (P < 0.05) higher than SLA or SLA plus ScLL 10 weeks after the challenge infection. The detection of high levels of IgG2a and the expression of mRNA cytokines, such as IFN-$\gamma$, IL-12, and TNF-$\alpha$ (Th1 profiles), corroborated the protective role of this lectin against cutaneous leishmaniasis. This is the first report of the ScLL effect on leishmaniasis and shows a promising role for ScLL to be explored in other experimental models for treatment of leishmaniasis.

• Fisheries and Aquatic Sciences
• /
• 제15권1호
• /
• pp.43-47
• /
• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.

• 생명과학회지
• /
• 제20권9호
• /
• pp.1420-1425
• /
• 2010

선천성 면역이란 감염성 질환에 대응하는 분자들의 네트워크가 반응하는 숙주의 첫 번째 방어 메카니즘이다. 간에서 만들어져 혈액에 존재하는 Mannose-binding lectin (MBL)은 선천성 면역에 관여하는 단백질군인 collectin에 속하는 분자로서 감염성 질병을 유발하는 다양한 세균, 바이러스, 효모, 곰팡이 및 원생동물의 표면에 존재하는 특징적인 당쇄를 인식한다. 이런 감염성 인자들의 표면에 드러난 당쇄의 공통적인 패턴을 MBL이 인식하여 자기(self)와 비자기(non-self)를 구분하기 때문에 MBL을 패턴 인식 분자(pattern recognition molecule)라고 한다. MBL은 MBL2 유전자에 의해 만들어지며, MBL2 유전형은 여러 가지 다형성(polymorphisms)이 있는 것으로 나타났다. MBL2 유전자의 변이는 상당히 많은 사람에서 나타나며, MBL 결여의 원인이다. MBL 결여는 감염성 질환에 대한 감수성을 증가시키므로, MBL의 유전적 변이와 임상적 중요성에 대해 많은 연구가 진행 되어져 왔다. 이 총설은 현재 우리가 알고 있는 MBL의 구조와 기능에 대해 전반적으로 논의하고자 한다.