A Low-molecular-weight Inhibitor of the Neutral Proteinase from Rat Intestinal Smooth Muscle

Overview

Journal Biochem J

Specialty Biochemistry

Date 1980 Feb 1

PMID 7396824

Citations 2

Authors

I T Carney

C G Curtis

J K Kay

N Birket

Affiliations

Soon will be listed here.

Abstract

1. Rat intestinal smooth muscle was shown to contain endogenous inhibitory activity towards the neutral trypsin-like muscle proteinase described previously [Beynon & Kay (1978) Biochem. J. 173, 291--298]. 2. Comtamination of the muscle tissue by mucosal, blood and pancreatic inhibitors was shown to be unlikely. 3. The inhibitory activity was resolved into high- and low-molecular-weight components. 4. The low-molecular-weight component was purified to homogeneity. It has a molecular weight of approx. 9000 and was stable over the pH range 3--11. 5. It inhibited the muscle proteinase competitively (Ki congruent to t microM), but had no effect on any of the other proteinases tested. 6. Leupeptin also inhibited the muscle proteinase competitively (Ki congruent to 0.3 microM), whereas the low-molecular weight proteins gastrin, glucagon and insulin B-chain had very little effect. 7. A role for a weakly binding inhibitor in modulating the influence of the neutral proteinase on intracellular protein degradation is considered.

Citing Articles

Degradation of myofibrillar proteins by trypsin-like serine proteinases.

Kay J, Siemankowski L, Siemankowski R, Greweling J, Goll D Biochem J. 1982; 201(2):279-85.

PMID: 7044373 PMC: 1163641. DOI: 10.1042/bj2010279.

Degradation of smooth-muscle myosin by trypsin-like serine proteinases.

Kay J, Siemankowski R, Siemankowski L, Goll D Biochem J. 1982; 201(2):267-78.

PMID: 6123314 PMC: 1163640. DOI: 10.1042/bj2010267.

References

Greene L, Rigbi M, Fackre D . Trypsin inhibitor from bovine pancreatic juice. J Biol Chem. 1966; 241(23):5610-8. View

Schroeder D, Shaw E . Chromatography of trypsin and its derivatives. Characterization of a new active form of bovine trypsin. J Biol Chem. 1968; 243(11):2943-9. View

PUTTER J . [On the kinetics of the reaction between kallikrein inactivator and trypsin]. Hoppe Seylers Z Physiol Chem. 1967; 348(9):1197-206. View

Weber K, Osborn M . The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969; 244(16):4406-12. View

Swank R, MUNKRES K . Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971; 39(2):462-77. DOI: 10.1016/0003-2697(71)90436-2. View

Kay J, Kassell B . The autoactivation of trypsinogen. J Biol Chem. 1971; 246(21):6661-5. View

Katunuma N . Regulation of intracellular enzyme levels by limited proteolysis. Rev Physiol Biochem Pharmacol. 1975; 72:83-104. DOI: 10.1007/BFb0031547. View

Dunaway G, SEGAL H . Purification and physiological role of a peptide stabilizing factor of rat liver phosphofructokinase. J Biol Chem. 1976; 251(8):2323-9. View

Knowles S, Ballard F . Selective control of the degradation of normal and aberrant proteins in Reuber H35 hepatoma cells. Biochem J. 1976; 156(3):609-17. PMC: 1163795. DOI: 10.1042/bj1560609. View

10.

KIRSCHKE H, Langner J, Wiederanders B, Ansorge S, Bohley P . Cathepsin L. A new proteinase from rat-liver lysosomes. Eur J Biochem. 1977; 74(2):293-301. DOI: 10.1111/j.1432-1033.1977.tb11393.x. View

11.

Osterlund B, BRIDGER W . Effect of a peptide stabilizing factor on liver ATP citrate lyase. Biochem Biophys Res Commun. 1977; 76(1):1-8. DOI: 10.1016/0006-291x(77)91660-6. View

12.

Hopgood M, Clark M, Ballard F . Inhibition of protein degradation in isolated rat hepatocytes. Biochem J. 1977; 164(2):399-407. PMC: 1164805. DOI: 10.1042/bj1640399. View

13.

Ballard F . Intracellular protein degradation. Essays Biochem. 1977; 13:1-37. View

14.

Woodbury R, Gruzenski G, Lagunoff D . Immunofluorescent localization of a serine protease in rat small intestine. Proc Natl Acad Sci U S A. 1978; 75(6):2785-9. PMC: 392649. DOI: 10.1073/pnas.75.6.2785. View

15.

Carney I, Kay J, Birket N . An endogenous inhibitor of the neutral proteinase from rat intestinal smooth muscle [proceedings]. Biochem Soc Trans. 1978; 6(3):550-2. DOI: 10.1042/bst0060550. View

16.

Beynon R, Kay J . The inactivation of native enzymes by a neutral proteinase from rat intestinal muscle. Biochem J. 1978; 173(1):291-8. PMC: 1185773. DOI: 10.1042/bj1730291. View

17.

Beynon R, Kay J . A neutral protease from rat intestinal muscle. A possible role in the degradation of native enzymes. Acta Biol Med Ger. 1977; 36(11-12):1624-35. View

18.

Kay J, Carney I, Beynon R . The susceptibility of glycogen phosphorylase to inactivation by endogenous and exogenous proteases. Acta Biol Med Ger. 1977; 36(11-12):1637-44. View

19.

Bohley P, Riemann S, Koelsch R, Lasch J . Protein degradation in rat liver cells. Acta Biol Med Ger. 1977; 36(11-12):1821-2. View

20.

Dunn W, ARONSON Jr N . Inhibition of glycoprotein catabolism in vivo and in the perfused rat liver. Acta Biol Med Ger. 1977; 36(11-12):1917-21. View