Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new soluble 170-kDa protein (BP170) was found to be present exclusively in the brain of all the vertebrates that we studied by Western immunoblotting. It was not detected in peripheral rat tissues, including heart, kidney, liver, spleen, lung, muscle, adrenal, intestinal mucosa, sciatic nerve, or pituitary. In rat brain, its regional distribution was found to be heterogeneous, with its highest concentration in the cerebrum and its lowest in the hypothalamus, and 89% of it was in the post-microsomal fraction. BP170 constitutes at least 0.05% of the total brain cytosol proteins. Its level increases during development, being the lowest at 5 days and the highest at 90 days postnatal. BP170 is a single-chain polypeptide. It could be partially purified by precipitation with polyethylene glycol followed by column chromatography on Q Sepharose. Although BP170 was identified by an antiserum against puromycin-sensitive aminopeptidase (PSA), the two proteins differ in molecular weight, chromatographic properties, regional and subcellular distribution, developmental changes, immunoreactivity, and enzyme activity. Self-incubation or trypsin treatment of the partially purified BP170 generates no PSA activity, indicating that BP170 is not a PSA precursor. Furthermore, BP170 is neither an inhibitor nor an activator of PSA. Our data suggest that BP170 is a novel brain-specific protein not previously described.
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PMID:A new soluble brain-specific protein: identification and partial purification. 846 1

Two puromycin-sensitive aminopeptidase isozymes (PSA-I and PSA-II) were isolated from chicken brain cytosol by ammonium sulfate fractionation followed by column chromatography on Cellex D and AH-Sepharose 4B and separated on Bio-Gel HTP. Each was purified to homogeneity on Sephadex G-200, Arg-Tyr-AH-Sepharose, Bio-Gel HTP, and preparative gel electrophoresis. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, PSA-I appeared to be a monomer with a molecular mass of 105 kDa, and PSA-II to be composed of two subunits of 25 kDa and 100 kDa. The tryptic maps of 100 kDa and 105 kDa protein in HPLC are different in peak frequency, height, and composition. The internal peptide sequence of PSA-I has a considerable homology to PSA-II. Both isozymes have repeated copies of common peptide segments and have no significant sequence homology to other peptidases and proteinases. These thio and Co(2+)-activated isozymes have a neutral pH optimum and are inhibited by puromycin and bestatin. PSA-II is more sensitive to trypsin and heat treatment, has a lower Km to Met-enkephalin, and is more active on Arg BNA and Pro BNA. Our results suggest that PSA-I and PSA-II derive from translation of two RNAs of a new gene family related to the brain-specific 14-3-3 protein.
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PMID:Two cytosolic puromycin-sensitive aminopeptidase isozymes in chicken brain: molecular homology to brain-specific 14-3-3 protein. 848 50

The puromycin-sensitive aminopeptidase was found to be resistant to proteolysis by trypsin, chymotrypsin, and protease V8 but was cleaved into an N-terminal 60-kDa fragment and a C-terminal 33-kDa fragment by proteinase K. The two proteinase K fragments remain associated and retained enzymatic activity. Attempts to express the 60-kDa N-terminal fragment in Escherichia coli produced inclusion bodies. A hexa-histidine fusion protein of the 60-kDa N-terminal fragment was solubilized from inclusion bodies with urea and refolded by removal of the urea through dialysis. The refolded protein was devoid of aminopeptidase activity as assayed with arginine-beta-naphthylamide. However, the refolded protein bound the substrate dynorphin A(1-9) with a stoichiometry of 0.5 mol/mol and a K(0.5) value of 50 microM. Dynorphin A(1-9) binding was competitively inhibited by the substrate dynorphin B(1-9), but not by des-Tyr(1)-leucine-enkephalin, a poor substrate for the enzyme.
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PMID:Proteolytic cleavage of the puromycin-sensitive aminopeptidase generates a substrate binding domain. 1280 15