Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.64 (proteinase K)
 4,071 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

X-linked ichthyosis is the result of steroid sulfatase (STS) deficiency. While most affected individuals have extensive deletions of the STS gene, point mutations have been reported in three patients (1). In this study, we identify an additional three point mutations and characterize the effects of all six mutations on STS activity and expression. All six are unique single base pair substitutions. The mutations are located in a 105-amino acid region of the C-terminal half of the polypeptide. Five of the six mutations involve the substitutions of Pro or Arg for Trp372, Arg for His444, Tyr for Cys446, or Leu for Cys341. The other mutation is in a splice junction and results in a frameshift causing premature termination of the polypeptide at residue 427. All the affected residues are conserved to some degree within the sulfatase family. The six mutations were reproduced in normal STS cDNA and transiently expressed in STS-deficient cells. All six mutant vectors direct the expression of STS protein that lacks enzymatic activity. The mutant polypeptides show a shift in mobility on SDS-PAGE and resistance to proteinase K digestion when translated in the presence of dog pancreas microsomes, indicating glycosylation and normal translocation.
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PMID:Characterization of point mutations in patients with X-linked ichthyosis. Effects on the structure and function of the steroid sulfatase protein. 925 98

As the cultivars of rice markedly affect eating quality, processing suitability, and price, identification or differentiation of rice cultivar is very important. We developed suitable 14 STS (sequence-tagged site) primers for PCR (polymerase chain reaction), and it became possible to differentiate 60 Japanese dominant rice cultivars from each other using template DNA extracted and purified from rice grains. A multiplex primer set was shown to be useful to effectively differentiate rice cultivars produced in various countries by PCR. A novel multiplex primer set for PCR has been developed to differentiate KoshihikariBL, which is closely related with the premium cultivar, Koshihikari, in Japan. The application of the cultivar identification method by PCR method to commercially processed rice products was investigated. We developed an enzyme treatment method, in which the gelatinized starch is decomposed by the heat-stable alpha-amylase at 80 degrees C, followed by the hydrolysis of proteins by proteinase K with sodium dodecyl sulfate and purification of extracted DNAs by phenol/chloroform/iso-amyl alcohol. It became possible to identify the material rice cultivars of the commercially processed rice products, such as cooked rice, rice cake, or rice cracker, by a PCR method using template DNA prepared by the enzyme treatment method and novel multiplex primer sets.
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PMID:Cultivar identification of rice (Oryza sativa L.) by polymerase chain reaction method and its application to processed rice products. 1725 60

We developed a method for the preparation of template DNAs for polymerase chain reaction (PCR) from beer. We improved the method by (i) lyophilizing and pulverizing the beer to concentrate DNAs, (ii) decomposition of polysaccharides and proteins so as not to inhibit DNA extraction by the use of heat-resistant amylase and proteinase K, (iii) separation of template DNA by purification using 70% EtOH extraction and isopropyl alcohol precipitation, and (iv) the use of magnetic beads to purify DNA. We developed suitable 7 STS (sequence-tagged site) primers related to beer quality for PCR, and it proved possible to identify 16 dominant malting barley cultivars and 22 kinds of beers. To digitize the results of PCR, discriminative DNA bands were binarized as 0 (disappeared) or 1 (appeared) and subjected to multiple regression analysis. Estimation formulae for the quality of beer were developed using the above-mentioned independent variables based on the results of PCR against dependent variables related to the qualities of beer, including foam stability, bitterness, sourness and astringency. These equations showed multiple regression coefficients of 0.93, 0.82, 0.87, and 0.87 for calibration.
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PMID:A novel method for the preparation of template DNA for PCR from beer to detect materials and to develop DNA markers to evaluate the quality of beer. 2356 68