Calpain from Rat Intestinal Epithelial Cells: Age-dependent Dynamics During Cell Differentiation

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 1994 Feb 9

PMID 8047065

Citations 3

Authors

M Ibrahim

R K Upreti

A M Kidwai

Affiliations

Soon will be listed here.

Abstract

Micromolar and millimolar Ca(2+)-requiring neutral protease (calpain I and calpain II) along with their endogenous inhibitor calpastatin were isolated and partially purified from the same preparation of rat intestinal epithelial cells. Calpain I and II were partially purified by 1300 and 900-fold with 57 and 53 per cent yield, respectively. The optimum assay conditions revealed pH 7.5, 20 min incubation at 25 degrees C and 0.24% casein substrate for both calpains. The optimum calcium concentration obtained for calpain I and II were 25 microM and 4 mM, respectively. Distribution of rat intestinal epithelial cells calpain I and II along with calpastatin during cell differentiation stages in weanling to senescence age were studied. Calpain I in weanling rats was in an increasing order from villus to crypt regions. Adult rats indicated well expressed consistent calpain I throughout the differentiation stages. Whereas, significant lowering towards crypt region cells were evident in old rats. Calpain II in weanling and adult rats was found to be consistent throughout the differentiation stages. Old animals revealed an increasing trend from villus to crypt region with insignificant activity present in upper villus cells. Concomitantly, different concentrations of calpastatin were observed throughout the differentiation stages in all the age groups. Moreover, the levels of calpains exceeded that of calpastatin in most of the epithelial cell populations during developmental stages. In addition to casein, intestinal epithelial cell membranes were found to be equally good substrates for calpains. Proteolytic susceptibility of weanling, adult and old rat membrane proteins varied significantly all along the ageing process in rats. Simultaneous age-dependent calpastatin response were also evident. Taken together the results obtained provided strong evidence that calpain plays significant role in rat intestinal cell differentiation and ageing process with calpastatin as its specific regulatory protein.

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References

SIMONSON L, Baudry M, Siman R, Lynch G . Regional distribution of soluble calcium activated proteinase activity in neonatal and adult rat brain. Brain Res. 1985; 327(1-2):153-9. DOI: 10.1016/0006-8993(85)91509-4. View

Kenessey A, Banay-Schwartz M, Deguzman T, Lajtha A . Calpain II activity and calpastatin content in brain regions of 3- and 24-month-old rats. Neurochem Res. 1990; 15(3):243-9. DOI: 10.1007/BF00968667. View

Waxman L . Calcium-activated proteases in mammalian tissues. Methods Enzymol. 1981; 80 Pt C:664-80. DOI: 10.1016/s0076-6879(81)80051-1. View

Schafer D, HULTQUIST D . Isolation of an acid protease from rabbit reticulocytes and evidence for its role in processing redox proteins during erythroid maturation. Biochem Biophys Res Commun. 1981; 100(4):1555-61. DOI: 10.1016/0006-291x(81)90696-3. View

Nixon R, Quackenbush R, Vitto A . Multiple calcium-activated neutral proteinases (CANP) in mouse retinal ganglion cell neurons: specificities for endogenous neuronal substrates and comparison to purified brain CANP. J Neurosci. 1986; 6(5):1252-63. PMC: 6568565. View

Kubota S, OHSAWA N, Takaku F . Purification of a calcium-activated neutral proteinase from human placenta. Biochim Biophys Acta. 1984; 802(2):379-83. DOI: 10.1016/0304-4165(84)90186-7. View

Yoshimura N, Tsukahara I, Murachi T . Calpain and calpastatin in porcine retina. Identification and action on microtubule-associated proteins. Biochem J. 1984; 223(1):47-51. PMC: 1144262. DOI: 10.1042/bj2230047. View

FORSTNER G, SABESIN S, Isselbacher K . Rat intestinal microvillus membranes. Purification and biochemical characterization. Biochem J. 1968; 106(2):381-90. PMC: 1198514. DOI: 10.1042/bj1060381. View

SCHLAEPFER W, Lee C, Lee V, Zimmerman U . An immunoblot study of neurofilament degradation in situ and during calcium-activated proteolysis. J Neurochem. 1985; 44(2):502-9. DOI: 10.1111/j.1471-4159.1985.tb05442.x. View

10.

Dayton W, Goll D, Zeece M, Robson R, Reville W . A Ca2+-activated protease possibly involved in myofibrillar protein turnover. Purification from porcine muscle. Biochemistry. 1976; 15(10):2150-8. DOI: 10.1021/bi00655a019. View

11.

Gilchrist J, Wang K, Katz S, Belcastro A . Calcium-activated neutral protease effects upon skeletal muscle sarcoplasmic reticulum protein structure and calcium release. J Biol Chem. 1992; 267(29):20857-65. View

12.

Siman R, Baudry M, Lynch G . Brain fodrin: substrate for calpain I, an endogenous calcium-activated protease. Proc Natl Acad Sci U S A. 1984; 81(11):3572-6. PMC: 345551. DOI: 10.1073/pnas.81.11.3572. View

13.

Nixon R . Calcium-activated neutral proteinases as regulators of cellular function. Implications for Alzheimer's disease pathogenesis. Ann N Y Acad Sci. 1989; 568:198-208. DOI: 10.1111/j.1749-6632.1989.tb12509.x. View

14.

Siman R, Noszek J . Excitatory amino acids activate calpain I and induce structural protein breakdown in vivo. Neuron. 1988; 1(4):279-87. DOI: 10.1016/0896-6273(88)90076-1. View

15.

Miyachi Y, Yoshimura N, Suzuki S, Hamakubo T, Kannagi R, Imamura S . Biochemical demonstration and immunohistochemical localization of calpain in human skin. J Invest Dermatol. 1986; 86(4):346-9. DOI: 10.1111/1523-1747.ep12285556. View

16.

Gaczynska M . Changes in the proteolytic activity of human erythrocyte membrane during red cell aging. Biochim Biophys Acta. 1989; 981(2):173-7. DOI: 10.1016/0005-2736(89)90025-4. View

17.

Farber J . The role of calcium ions in toxic cell injury. Environ Health Perspect. 1990; 84:107-11. PMC: 1567653. DOI: 10.1289/ehp.9084107. View

18.

Kirkland J . The biochemistry of mammalian senescence. Clin Biochem. 1992; 25(2):61-75. DOI: 10.1016/0009-9120(92)80047-k. View

19.

Oshima M, Koizumi S, Fujita K, Guroff G . Nerve growth factor-induced decrease in the calpain activity of PC12 cells. J Biol Chem. 1989; 264(34):20811-6. View

20.

Ashby C, Walsh D . Purification and characterization of an inhibitor protein of cyclic AMP-dependent protein kinases. Methods Enzymol. 1974; 38:350-8. DOI: 10.1016/0076-6879(74)38051-2. View