PMID: 7023544Jul 24, 1981Paper

Purification and characterization of alkaline protease and neutral protease from chromatin of rats

Biochimica Et Biophysica Acta
H HagiwaraT Horio

Abstract

It was previously reported that, in addition to a known chymotrypsin-like protease capable of hydrolyzing histones with an optimum pH of 8 (neutral protease), another protease is bound to the chromatin of various rat tissues and in situ hydrolyzes casein more quickly than histones with an optimum pH of 10 (alkaline protease). In the present study, the alkaline protease was purified 14 000-fold to approx. 75% purity from the chromatin of Rhodamine sarcoma. This tumor contains both proteases at higher levels than normal tissues. For purification, affinity columns of Sepharose with bound soybean trypsin inhibitor, casein and histones were successively used. Also, the neutral protease was purified 920-fold to an apparently homogeneous state by affinity chromatography on a Sepharose column with bound soybean trypsin inhibitor under conditions, in which an excess amount of the enzyme was applied on the column so that part of the enzyme would pass through the column without adsorption and the enzyme thus adsorbed was then eluted. The purified alkaline and neutral proteases had molecular weights of approx. 18 000 and 27 000, respectively, and isoelectric points of approx. 11. The former enzyme hydrolyzed casein (100) in preference to h...Continue Reading

References

Dec 1, 1976·Archives of Biochemistry and Biophysics·M JusicP C Heinrich
Jul 1, 1977·Journal of Biochemistry·Y Suzuki, T Murachi
Jan 24, 1975·Nature·J P BaldwinK Ibel
Aug 27, 1968·Biochimica Et Biophysica Acta·M FurlanA Suhar
Mar 1, 1974·Science·G S SteinL J Kleinsmith
Sep 1, 1973·Archives of Biochemistry and Biophysics·J F Dice, R T Schimke
Mar 28, 1969·Journal of Molecular Biology·R Hancock
Jun 16, 1980·Biochemical and Biophysical Research Communications·H HagiwaraT Horio
Feb 1, 1961·Archives of Biochemistry and Biophysics·N K SARKAR, A L DOUNCE

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Citations

Oct 29, 2014·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Maarten DhaenensDieter Deforce
Nov 15, 1985·Biochemical and Biophysical Research Communications·N YamashitaT Horio
Dec 8, 1989·European Journal of Biochemistry·S MiyataH K Kihara
Nov 22, 1983·Biochimica Et Biophysica Acta·Y Marushige, K Marushige
Jul 16, 1982·Biochemical and Biophysical Research Communications·Y MatuoF Wada
Nov 30, 1982·Biochemical and Biophysical Research Communications·Y MatuoF Wada

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