• BMB Reports
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• 제30권5호
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• pp.356-361
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• 1997

We cloned a cDNA (pPRC-1) which was comprised of 841 nucleotides from the cDNA library of a male ICR mouse submandibular gland ($SMG^+$). The nucleotide sequences of pPRC-1 were identical to those of exons 2 and 3 of the mGK-21 gene, a potentially functional glandular kallikrein identified in a Balb/c mouse, except for one nucleotide residue. Although this substitution changes Ile (ATT) in pPRC-1 to Val (GTT) in mGK-21, this difference has been explained by strain polymorphism. From the amino acid sequences predicted from its cDNA, we speculated that mGK-21 gene products/pGK21 consist of 261 amino acids including the $NH_2$-terminal signal peptide (residues 1~17), the short propeptide (residues 17~24), and the active peptide (residues 25~261). Although we did not demonstrate the enzyme activity of pGK21, it was assumed that pGK 21 was involved in the maturation of certain bioactive polypeptide(s) in mouse SMG for the following reasons : (a) mGK-21 gene was apparently expressed in a male ICR mouse SMG: (b) the proposed active site $His^{65}$, $Asp^{120}$, and $Ser^{213}$ residues were completely conserved in pGK21 just like other glandular kallikreins; (c) the cloned cDNA was translated to a predicted 27 kDa polypeptide chain in vitro: (d) the 27 kDa polypeptide chain produced by CHO cells was produced to a putative active form by trypsin.

• 한국동물학회지
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• 제39권2호
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• pp.215-222
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• 1996

In the previous studies, we have demonstrated that prorenin converting enzyme (PRECE) was identical to the epidermal grouch factor-binding protein (EGF-BP) type B, which was a member of the mouse glandular kallikrein family, To examine whether or not EGF-BP type A and C are involved in the processing of prorenin, we have cloned the CDNAS of the EGF-BP type h and C from a library of male ICR mouse submandibular gland (SMGI. And then CHO cells were transfected with the EGF-BP expression plasmids. and stable cell lines expressing a high level of the EGF-BPS precursor were obtained. The conditioned medium was then treated with trypsin, which has been knotvn to effectively convert the EGF-BP type A and C precursor to the active forms. 수ubsequentlv, the prorenin converting activity of the trypsin-treated or untreated medium was examined. PRECE converted exactly prorenin to renin, but the prorenin converting activities of EGF-BP type A and C were not detected. From these results, it seems that only type B of these EGF-BPs is involved in processing Ren 2 prorenin in mouse SMG.

• KSBB Journal
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• 제24권1호
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• pp.80-88
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• 2009

glandular kallikrein에 광범위한 상동성을 가지는 인간 전립성 산성 인산 가수분해 효소는, 전립선암의 대표적인 혈청 biomarkers이다. 수지상세포는 유력한 항원 제시 세포이며, 바이러스, 미생물 병원체 및 종양에 대하여 면역 계통에서 강력한 T 세포 응답을 유도할 수 있다. 따라서, 종양 특이적인 항원으로 감작된 수지상세포를 이용한 면역요법은 anti-tumor 면역 유도를 위한 강력한 방법중의 하나이다. 크레아젠(주)에서 개발된 CTP (세포막 투과성 펩티드) 기술은 세포 내로의 높은 침투 효율성을 가지며 핵산이나 단백질과 같은 생체 고분자 물질을 접합하여 세포내로 수송할 수 있는 기술이다(36). 하지만 활성형의 인간 전립성 산성 인산 가수분해 효소는 세포사멸을 매개할 수 있기 때문에, 본 연구진은 항암 치료용 백신으로의 수지상세포 감작을 위해 비활성형 형태의 다중체 (multimer) 항원을 개발하였다. 본 연구에서, 수지상 세포의 감작과 활성화에 안전하고 효율적인 다중체 형태 (multimeric form)의 세포막 투과성 펩티드가 융합된 인간 전립성 산성 인산 가수분해 효소를 얻기 위한 정제공정을 기법을 개발하였고 젤 여과 크로마토그래피, western blot과 Dynamic Light Scattering을 이용하여 확인하였다. 아울러, 정제된 다중체 형태 (multimeric form)의 세포막 투과성 펩티드가 융합된 인간 전립성 산성 인산 가수분해 효소는 수지상 세포의 감작시 세포질 내로 효과적으로 침투되었다. 결과적으로 세포의 사멸의 부작용이 없이 MHC class I 분자를 통해 수지상세포의 표면으로 효과적으로 제시되었다.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.840-846
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• 2007

Human prostate-specific antigen(PSA), a 33 kDa serine protease with comprehensive homology to glandular kallikrein, is secreted from prostatic tissue into the seminal fluid and enters into the circulation. The level of PSA increases in the serum of patients with prostatic cancer and hence is widely employed as a marker of the disease status. In particular, an enzymatically active PSA that is a form cleaved at the N-terminal seven-amino-acids prosequence, APLILSR, of proPSA may play an important roll in the progression of prostate cancer. Thus, the presence of the active form would selectively discriminate the cancer from benign prostatic hyperplasia. In this study, we developed a convenient purification method for the acquisition of active PSA and proPSA. Recombinant proPSA and active PSA were expressed directly in Escherichia coli, easily and efficiently isolated from inclusion bodies, refolded, and purified. Moreover, the enzymatic activity of the recombinant active PSA was confirmed as serine protease using chromogenic chymotrypsin substrate. This purified active PSA could be further applied to scrutinize the biological or conformational characteristics of the protein and to develop specific diagnostic and/or therapeutic agents against prostate cancer.