Effect of selected Hofmeister cations and anions on recombinant protease B solubility

2012 ◽  
Vol 1 (2) ◽  
Author(s):  
Sk. Amir Hossain ◽  
Andres Veide ◽  
Cecilia Janson Kapca ◽  
Kim Hanson Uhre ◽  
Sune Jakobsen
Keyword(s):  
Protease B

Protease B of Saccharomyces cerevisiae: Isolation and Regulation of the PRB1 Structural Gene

Genetics ◽  
1987 ◽  
Vol 115 (2) ◽  
pp. 255-263 ◽  
Author(s):  
Charles M Moehle ◽  
Martha W Aynardi ◽  
Michael R Kolodny ◽  
Frances J Park ◽  
Elizabeth W Jones
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Weight ◽  
Structural Gene ◽  
Genomic Library ◽  
Time Lag ◽  
Proteolytic Processing ◽  
Coding Region ◽  
Protease B ◽  
Endonuclease Mapping ◽  
Protein Molecular Weight

ABSTRACT We have isolated the structural gene, PRB1, for the vacuolar protease B of Saccharomyces cerevisiae from a genomic library by complementation of the prb1-1122 mutation. Deletion analysis localized the complementing activity to a 3.2-kilobase pair XhoI-HindIII restriction enzyme fragment. The fragment was used to identify a 2.3-kilobase mRNA. S1 endonuclease mapping indicated that the mRNA and the gene were colinear. No introns were detected. The mRNA is of sufficient size to encode a protein of about 69,000 molecular weight, a number much larger than either the mature enzyme (≃30,000 protein molecular weight) or the sole reported precursor (≃39,000 protein molecular weight). These results suggest that proteolytic processing steps beyond that thought to be catalyzed by protease A may be required to convert the initial glycosylated translation product into mature protease B. The PRB1 mRNA is made in substantial amounts only when the cells have exhausted the glucose supply and enter the diauxic plateau. There is an extended time lag between PRB1 transcription and expression of protease B activity. A deletion that removes about 83% of the coding region was constructed as a diploid heterozygote. Spores bearing the deletion germinate, grow at normal rates into colonies, and have no obvious phenotype beyond protease B deficiency.

scholarly journals The PBN1 Gene of Saccharomyces cerevisiae: An Essential Gene That Is Required for the Post-translational Processing of the Protease B Precursor

Genetics ◽  
1998 ◽  
Vol 149 (3) ◽  
pp. 1277-1292 ◽  
Author(s):  
Rajesh R Naik ◽  
Elizabeth W Jones
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Secretory Pathway ◽  
Transmembrane Domain ◽  
Essential Gene ◽  
Signal Sequence ◽  
Signal Peptidase ◽  
Amino Terminal ◽  
Autocatalytic Cleavage ◽  
Protease B ◽  
Chromosome Iii

Abstract The vacuolar hydrolase protease B in Saccharomyces cerevisiae is synthesized as an inactive precursor (Prb1p). The precursor undergoes post-translational modifications while transiting the secretory pathway. In addition to N- and O -linked glycosylations, four proteolytic cleavages occur during the maturation of Prb1p. Removal of the signal peptide by signal peptidase and the autocatalytic cleavage of the large aminoterminal propeptide occur in the endoplasmic reticulum (ER). Two carboxy-terminal cleavages of the post regions occur in the vacuole: the first cleavage is catalyzed by protease A and the second results from autocatalysis. We have isolated a mutant, pbn1-1, that exhibits a defect in the ER processing of Prb1p. The autocatalytic cleavage of the propeptide from Prb1p does not occur and Prb1p is rapidly degraded in the cytosol. PBN1 was cloned and is identical to YCL052c on chromosome III. PBN1 is an essential gene that encodes a novel protein. Pbn1p is predicted to contain a sub-C-terminal transmembrane domain but no signal sequence. A functional HA epitope-tagged Pbn1p fusion localizes to the ER. Pbn1p is N-glycosylated in its amino-terminal domain, indicating a lumenal orientation despite the lack of a signal sequence. Based on these results, we propose that one of the functions of Pbn1p is to aid in the autocatalytic processing of Prb1p.

scholarly journals THE MECHANISM OF ANAPHYLACTIC SHOCK

1915 ◽  
Vol 22 (4) ◽  
pp. 401-417 ◽  
Author(s):  
James W. Jobling ◽  
William Petersen ◽  
A. A. Eggstein
Keyword(s):  
Amino Acids ◽  
Anaphylactic Shock ◽  
Serum Proteins ◽  
Progressive Increase ◽  
Acute Intoxication ◽  
Foreign Protein ◽  
Serum Lipase ◽  
Maximum Period ◽  
Specific Protease ◽  
Protease B

1. The serum ferments are practically unaltered by a primary injection of foreign protein. 2. During the course of sensitization the injection of the antigen is followed by the mobilization of a non-specific protease which increases in rapidity and intensity as the maximum period of sensitization is reached. 3. Acute shock is accompanied by: (a) The instantaneous mobilization of a large amount of non-specific protease; (b) a decrease in antiferment; (c) an increase in non-coagulable nitrogen of the serum; (d) an increase in amino-acids; (e) a primary decrease in serum proteoses. 4. Later there is a progressive increase in the non-coagulable nitrogen, in proteoses, and in serum lipase. 5. The acute intoxication is brought about by the cleavage of serum proteins (and proteoses) through the peptone stage by a non-specific protease. 6. The specific elements lie in the rapid, mobilization of this ferment and the colloidal serum changes which bring about the change in antiferment titer.

Structure of the complex of Streptomyces griseus protease B and the third domain of the turkey ovomucoid inhibitor at 1.8-.ANG. resolution

Biochemistry ◽  
1983 ◽  
Vol 22 (19) ◽  
pp. 4420-4433 ◽  
Author(s):  
Randy J. Read ◽  
Masao Fujinaga ◽  
Anita R. Sielecki ◽  
Michael N. G. James
Keyword(s):  
Streptomyces Griseus ◽  
The Third ◽  
Protease B

scholarly journals Active-site variants of Streptomyces griseus protease B with peptide-ligation activity

2000 ◽  
Vol 7 (3) ◽  
pp. 163-171 ◽  
Author(s):  
R Jeffery Elliott ◽  
Andrew J Bennet ◽  
Curtis A Braun ◽  
Alasdair M MacLeod ◽  
Thor J Borgford
Keyword(s):  
Active Site ◽  
Streptomyces Griseus ◽  
Protease B ◽  
Peptide Ligation

scholarly journals Inactivation of Scytalidium lignicolum acid protease B with 1,2-epoxy-3-(4'-azido-2'-nitrophenoxy)propane.

1989 ◽  
Vol 53 (10) ◽  
pp. 2751-2756 ◽  
Author(s):  
Daisuke TSURU ◽  
Akihiko NAOTSUKA ◽  
Ryuji KOBAYASHI ◽  
Tadashi YOSHIMOTO ◽  
Kohei ODA ◽  
...  
Keyword(s):  
Acid Protease ◽  
Protease B

Isolation and characterization of yeast protease B

1979 ◽  
Vol 86 (3) ◽  
pp. 815-821 ◽  
Author(s):  
Yukihiro Sanada ◽  
Katsuhisa Fujishiro ◽  
Hideshi Tanaka ◽  
Nobuhiko Katunuma
Keyword(s):  
Isolation And Characterization ◽  
Protease B
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