• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2665-2668
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• 2014

Aptamers are oligonucleotides or peptide molecules that are able to bind to their specific target molecules with high affinity via molecular recognition. In this study, we present development of aptamer-based precipitation assays (or simply aptamoprecipitation) for His-tagged proteins and thrombin to compare their purification efficiency with other conventional affinity precipitation methods. A crosslinking method was employed to immobilize thiol-functionalized aptamers onto the surface of polystyrene resins, enabling them to specifically bind to His-tag and to thrombin, respectively. The resulting aptamer-functionalized resins were successfully applied via a one-step experiment to purification of His-tagged proteins from complex E. coli and to thrombin extraction, exhibiting superior or at least comparable purification results to the conventional immobilized metal affinity precipitation or immunoprecipitation.

• BMB Reports
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• 제49권6호
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• pp.319-324
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• 2016

RNA polymerase II C-terminal domain phosphatases are newly emerging family of phosphatases that contain FCPH domain with Mg⁺²-binding DXDX(T/V) signature motif. Its subfamily includes small CTD phosphatases (SCPs). Recently, we identified several interacting partners of human SCP1 with appearance of dephosphorylation and O-GlcNAcylation. In this study, using an established cell line with inducible CTDSPL2 protein (a member of the new phosphatase family), proteomic screening was conducted to identify binding partners of CTDSPL2 in nuclear extract through immunoprecipitation of CTDSPL2 with its associated. This approach led to the identification of several interacting partners of CTDSPL2. This will provide a better understanding on CTDSPL2.

• Journal of Microbiology and Biotechnology
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• 제1권4호
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• pp.232-239
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• 1991

Caboxymethyl cellulase (CMCase) I was purified from the induced culture filtrate of Penicllium verruculosum F-3 by ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Bio-gel P-150 filtration. The purified enzyme was assumed to be a glycoprotein consisting of 8.5% carbohydrate and having a molecular weight of 70.000 in SDS-polycrylamide gel electrophoresis (SDS-PAGE). The purified enzyme-specific anti-CMCase I IgG was obtained by rabbit immunization and protein A-sepharose CL-4B chromatography. The fungal poly($A^+$) RNA was isolated from the total RNA of the mycelium grown under cellulase induction conditions by oligo(dT)-cellulosse chromatography. The translation products in vitro were prepared by translating the isolated poly ($A^+$) RNA in rabbit reticulocyte lysate and analyzed by SDS-PAGE and fluorography. Of the translation products, CMCase I was identified by the immunoprecipitation against anti-CMCase I IgG.

• Genomics & Informatics
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• 제12권4호
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• pp.145-150
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• 2014

Recent technical advances, such as chromatin immunoprecipitation combined with DNA microarrays (ChIp-chip) and chromatin immunoprecipitation-sequencing (ChIP-seq), have generated large quantities of high-throughput data. Considering that epigenomic datasets are arranged over chromosomes, their analysis must account for spatial or temporal characteristics. In that sense, simple clustering or classification methodologies are inadequate for the analysis of multi-track ChIP-chip or ChIP-seq data. Approaches that are based on hidden Markov models (HMMs) can integrate dependencies between directly adjacent measurements in the genome. Here, we review three HMM-based studies that have contributed to epigenetic research, from a computational perspective. We also give a brief tutorial on HMM modelling-targeted at bioinformaticians who are new to the field.

• 약학회지
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• 제50권4호
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• pp.263-267
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• 2006

CD1d is an unique antigen presenting molecule which provides antigenic repertoires to NKT cells. To examine molecules required for CD1d antigen presentation, we determined an interaction between CD1d and several endoplasmic reticulum (ER) resident molecular chaperones by co-immunoprecipitation. Results indicated that calnexin and calreticulin seem to be bound to mouse CD1d, but TAP and tapasin do not bind. Further, we screened an yeat two hybrid system to identify proteins that help mouse CD1d transportation in the cytosol. We found that two proteins, heat shock protein a sub-unit $(Hsp90{\alpha})$ and protein kinase C and casein kinase substrate in neurons 3 (PACSIN-3), interact with CD1d. Future study will be focus on the role of these molecules during the CD1d antigen presentation.

• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.37-42
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• 1993

질트리코모나스(Trichomonasvqqin is)의 항원성 변이를 관찰하기 위해 원충의 표면 항원 (surface antigen)을 N-hydmwsuccinlmide-biotin(N반5-biotin)으로 표지 하고 표지된 표면 단백질과 토끼의 항혈청으로 면역침전(Immunoprecipitation; IP)시켰으며 sodium dodec낀 sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)와 전기영동이적법을 시 행하였다. 살아있는 원충을 NHS-biotin으로 표지하여 표면 단백질을 분리하고 이를 질트리코모나스에 면역된 토끼 항혈청과 면역침전 시켰던 바 46, 60, 68, 90, 130 그리고 220 kDa에서 6개의 단백질이 항원성을 나타내었으며 질토리코모나스의 분리주인 HY-1, HV-15 및 ATCC 50148 주간에 차이는 없어 이들 6개 분획이 표면 항원성 발현에 중요한 역할을 할 것으로 생각된다.

• 한국미생물·생명공학회지
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• 제33권3호
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• pp.189-193
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• 2005

RBL-2H3 cell lysates에 anti-protein phosphatase(PP) 1, 2A, 2B 항체를 첨가한 후 immunoprecipitation을 실시한 결과 PP2B를 가해준 샘플에서만 HRF를 확인하였다. 역으로 monoclonal anti-HRF 항체를 가한 후 immunoprecipitation을 실시한 결과 PP1, 2A는 검출되지 않았으나 PP2B의 경우는 regulatory subunit(19 kDa), catalyic subunit(60 kDa) 모두 확인할 수 있었다. Affinity chromatography를 통해서도 PP2B가 HRF의 탈인산화에 관여함을 확인하였다 즉 19kDa의 PP2B regulatory subunit과 60kDa의 catalytic subunit 모두가 확인되었으며 외부 $Ca^{2+}$이온 첨가 여부에 따른 차이는 관찰할 수 없었다. 결론적으로 RBL-2H3 cell에서 PP2B는 PP1이나 PP2A에 비해 상대적으로 그 존재량은 적으나 HRF와 상호작용하는 phosphatase로서 검출된 반면 PP1이나 PP2A는 검출되지 않았다.

• 대한수의학회지
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• 제32권1호
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• pp.83-90
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• 1992

소 설사병 바이러스 구조단백에 대한 단크론항체를 작성하여 혈청중화시험, 전기영동, 면역침전반응을 이용하여 분석하였던 바 다음의 결과를 얻었다. 중화능력이 있는 항체의 경우 56K내지 54K의 구조단백에 대응하였다. 그외 중화력을 나타내지 않는 항체는 45K와 36K의 바이러스 항원과 대응하였다. 순수정제된 바이러스의 전기영동 분석결과 12종 이상의 바이러스 단백성분이 구조단백질로서 검출되었으며 중화능력을 나타내는 항체를 이용한 면역침전 결과는 이들의 존재를 뒷받침하였다. 중화단백성분의 세포내 전구물질의 검출은 불가능하였으나 방사선동위원소 부착즉시 세포배지에서 바이러스의 존재를 확인할 수 있었다. Staphylococcus aureus $V_8$효소를 이용한 항원의 부분소화 분석결과 45K와 36K의 바이러스 항원은 서로 상관이 있는 것으로서 입증되었다.

• Journal of Microbiology and Biotechnology
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• 제32권7호
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• pp.938-948
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• 2022

Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

• Bulletin of the Korean Chemical Society
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• 제33권5호
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• pp.1765-1768
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• 2012