• Title/Summary/Keyword: proteinase K

Search Result 321, Processing Time 0.035 seconds

Isolation of Cysteine Proteinase Gene (PgCysP1) from Panax ginseng and Response of This Gene to Abiotic Stresses (인삼으로부터 Cysteine Proteinase 유전자의 분리 및 환경 스트레스에 대한 반응)

  • Jeong, Dae-Young;Kim, Yu-Jin;Shim, Ju-Sun;Lee, Jung-Hye;In, Jun-Gyo;Lee, Bum-Soo;Yang, Deok-Chun
    • Journal of Ginseng Research
    • /
    • v.32 no.4
    • /
    • pp.300-304
    • /
    • 2008
  • Cysteine proteinases play an essential role in plant growth and development but also in senescence and programmed cell death. They participate in both anabolic and catabolic processes. In addition, they are involved in signalling pathways and in the response to biotic and abiotic stresses. A cDNA clone encoding cysteine proteinase (CP) gene, designated PgCysP1, was isolated from Panax ginseng C. A. Meyer. Reverse transcriptase (RT)-PCR results showed that PgCysP1 expressed at different level in P. ginseng hairy root. Different stresses such as biotic as well as abiotic stresses triggered a significant induction of PgCysP1. The positive responses of PgCysP1 to the various stimuli suggested that PgCysP1 may help to protect the plant against reactive environmental stresses.

Characterization of 27K Zein as a Transmembrane Protein

  • Lee, Dong-Hee
    • BMB Reports
    • /
    • v.31 no.2
    • /
    • pp.196-200
    • /
    • 1998
  • Zeins, maize storage proteins, are retained in the endoplasmic reticulum (ER) during the subcellular targeting process without the ER retention signal. Circumstantial data indicate that the 27K zein is an ER transmembrane protein. The potential transmembrane domain may permit the 27K zein to remain in the ER. This study investigated the potential transmembrane feature by employing alkaline extraction, proteinase K digestion, and surface biotinylation on isolated intact protein bodies. These assays consistently support the possibility of the 27K zein as a transmembrane protein. The 27K zein polypeptide was shown to be associated with alkali-stripped membranes. The polypeptide was digested by proteinase K to a smaller fragment. According to surface biotinylation, the 27K zeins was labeled to the exclusion of other classes of zeins. This study, therefore, concludes that the 27K zein has an ER transmembrane domain, which may serve as an anchor for zeins' ER retention.

  • PDF

Study on Status of Nutritional Supply by Lunch-box in High School (고등학생(高等學生)의 도시락에 의한 영양섭취상태(營養攝取狀態)에 관(關)한 조사연구(調査硏究))

  • Rhee, Hei-Soo;Yim, Gong-Hee
    • Journal of Nutrition and Health
    • /
    • v.6 no.1
    • /
    • pp.39-46
    • /
    • 1973
  • This study was projected to get basic data which can provide a basis for future direction in nutritional education, and also to find the way how to improve the nutritional supply by evaluating the current nutritional intake of average high school students through the survey study of their daily packed lunch. Five hundred twenty seven students from two boys high school and two girls high school including one general and one vocational school respectively were chosen as random sampling technique. Four hundred forty nine among the 527 students had brought lunch. The contents of lunch box were weighed and converted into nutritional values according to the food composition table and compared with recommended dietary allowances. The results compared and classified by sex, School and housewives' educational level were as follows: 1. The nutritional supply in the lunch box was 671 Cal of energy and 22.3 gm of protein for male students which were respectively 55.9% and 74.2% of the dietary recommendations. On the other side female student's lunch boxes were found to contain 495 Cal of energy and 21.3gm of protein which are respectively 61.8% and 80% of the dietary prescriptions. Excluding niacin, all vitamins and minerals were found to be short. 2. Calorie intake in the vocational high school was found to be higher than in the general high school but lower in protein intake especially significant difference (P<0.01) in animal protein. 3. From the nutritional point of view the educational backgrouud of the housewives was not found to have any influence in the way of preparing the lunch boxes. 4. Nutrients of lunch box were heavily inclined to grain rather than to side dishes.

  • PDF

Effect of N-sources and NaCl Concentrations in Media on the Intra-and Extracellular Proteinase Activities of Streptococcus cremoris $ML_4$ and Streptococcus lactis $ML_8$ (Streptococcus cremoris $ML_4$ 및 Streptococcus lactis $ML_8$의 생육중 질소원과 염농도가 세포내 및 세포외 프로테이나제 역가에 미치는 영향)

  • Chang, Hae-Choon;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
    • /
    • v.21 no.1
    • /
    • pp.86-91
    • /
    • 1989
  • To investigate the effect of NaCl concentration and nitrogen sources in the medium for the Streptococci on the intra- and extracellular proteinase(ICP and ECP) which have been known as one of the major causes for the bitter peptide formation, Streptococcus lactis $ML_8$ and Streptococcus cremoris $ML_4$ strains were incubated at 0-4% NaCl in the medium and Na-caseinate as a nitrogen source 0-100%, and the cell growth, ICP and ECP activities were analyzed. As the concentration of the NaCl in the medium increased, the growth and ECP activity decreased but the ICP $activity/10^{10}$ cells increased on the contrary. This implied that the NaCl in the medium affects only the ECP which is associated mainly to the cell wall and cell membrane but not the ICP activity. When the content of the caseinate instead of other low molecular nitrogen sources were increased in the medium, the cell growth was lowered while the ECP activities increased probably by induction of proteinase production.

  • PDF

Production of Active Carboxypeptidase Y of Saccharomyces cerevisiae Secreted from Methylotrophic Yeast Pichia pastoris

  • RO, HYEON-SU;LEE, MI-SUN;HAHM, MOON-SUN;BAE, HEE-SUNG;CHUNG, BONG HYUN
    • Journal of Microbiology and Biotechnology
    • /
    • v.15 no.1
    • /
    • pp.202-205
    • /
    • 2005
  • Our previous study showed that the overexpression of carboxypeptidase Y (CPY) of Saccharomyces cerevisiae in Escherichia coli resulted in the formation of insoluble inclusion bodies. To produce soluble CPY, we designed a novel Pichia pastoris expression system, in which the following were inserted into expression vectors: three different signal sequences derived from the mating factor a1 of S. cerevisiae, an inulinase of Kluyveromyces marxianus, and the endogenous signal sequence of CPY. The expression vector pHIL-D2-SSinul-proCPY was the most effective in the production of proCPY among the vectors examined. The purified active CPY was obtained from proCPY by treating with proteinase K, followed by QExcellose ion-exchange column chromatography.

Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme

  • 나병국;송철용
    • Korean Journal of Microbiology
    • /
    • v.38 no.4
    • /
    • pp.218-218
    • /
    • 2002
  • Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme

  • Na, Byung-Kuk;Song, Chul-Yong
    • Journal of Microbiology
    • /
    • v.38 no.4
    • /
    • pp.218-223
    • /
    • 2000
  • Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

  • PDF

Isolation of the Gene for Lipocortin-1 Binding Protein Using Yeast Two Hybrid Assay (Yeast Two Hybrid Assay를 이용한 Lipocortin-1 결합 단백질 유전자의 분리)

  • Lee, Koung-Hoa;Kim, Jung-Woo
    • The Journal of Natural Sciences
    • /
    • v.9 no.1
    • /
    • pp.25-29
    • /
    • 1997
  • To study the mechanism of lipocortin-1, the 37 kDa protein, one of the annxin superfamily thought to be a second messenger during the Glucocorticoid dependent anti-inflammatory action, the gene for lipocortin-1 binding protein was isolated using the yeast two hybrid assay, the yeast based genetic assay recognizing the protein-protein interaction. The results showed that this gene has a weak homology to the for the human serine proteinase.

  • PDF

Sildenafil Citrate Induces Migration of Mouse Aortic Endothelial Cells and Proteinase Secretion

  • Kim, Young-Il;Oh, In-Suk;Park, Seung-Moon;Kim, Hwan-Gyu
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.11 no.5
    • /
    • pp.402-407
    • /
    • 2006
  • Vascular endothelial cells release proteinases that degrade the extracellular matrix (ECM), thus enabling cell migration during angiogenesis and vasculogenesis. Sildenafil citrate stimulates the nitric oxide-cyclic guanosine monophosphate pathway through inhibition of phosphodiesterase type V (PDE5). In this report, we examined the mechanisms underlying sildenafil citrate-induced cell migration using cultured mouse aortic endothelial cells (MAECs). Sildenafil citrate induced migration and proteinase secretion by murine endothelial cells. Sildenafil citrate induced the secretion of matrix metalloproteinase-2 (MMP-2) and MMP-9, which is inhibited by $NF-{\kappa}B$ inhibitors. Sildenafil citrate also induced the secretion of plasmin, which is inhibited by PI 3'-kinase inhibitors. It is suggested that sildenafil citrate-induced migrating activity in endothelial cells may be accomplished by increased secretion of proteinases.

Distribution and Structural Basis of the Native Strain in Human $\alpha_1$-Antitrypsin

  • Seo, Eun-Joo;Hana Im;Maeng, Jin-Soo;Kim, Kyoon-Eon;Yu, Myeong-Hee
    • Proceedings of the Korean Biophysical Society Conference
    • /
    • 1999.06a
    • /
    • pp.42-42
    • /
    • 1999
  • Metastability in the native form of proteins has been recognized as a mechanism of biological regulation. The strained native structure of serpins (serine proteinase inhibitors) is a typical example. The native strain of serpins is considered to be crucial to their physiological functions, such as plasma proteinase inhibition, hormone delivery, Alzheimer filament assembly, and extracellular matrix remodeling.(omitted)

  • PDF