Leucine Naphthylamide: an Appropriate Substrate for the Histochemical Detection of Cathepsins B and B′

Jk, McDonald; Bb, Zeitman; Ellis, Stanley

doi:10.1038/2251048a0

Cited by 59 publications

(38 citation statements)

References 12 publications

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“…Specifically, the present results are consistent with the occurrence of two soluble dipeptidylaminopeptidase activities, i.e. a dipeptidylaminopeptidase I11 with a pH optimum around 9.0 [25] and a second activity possibly classified as dipeptidylaminopeptidase I1 on the basis of substrate specificity [26,27] and pH optimum (6.7). The view of two distinct dipeptidylaminopeptidase activities in the soluble fraction of human erythrocytes receives further support by preliminary data indicating clearly differential patterns of decay of the activities toward the various substrates during aging of eryhthrocytes (unpublished observations).…”

Section: Discussionsupporting

confidence: 76%

Identification of Proteolytic Activities in the Cytosolic Compartment of Mature Human Erythrocytes

Pontremoli

Melloni

Salamino

et al. 1980

European Journal of Biochemistry

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Individual lysates from human erythrocyte suspensions, completely deprived of leucocytes and platelets, were assayed for a number of proteolytic activities using both naturally occurring and synthetic substrates. Removal of hemoglobin by batchwise DEAE-cellulose chromatography did not modify the complement of the various proteolytic activities which were then fractionated by means of chromatography on a column of DEAE-cellulose, followed by conventional techniques such as gel chromatography and preparative electrophoresis. This procedure allowed a number of proteinases to be identified in the erythrocyte cytosol while providing a tool for their selective though partial separation. The following peptidases were found to be present in the soluble fraction of mature human erythrocytes : (a) a neutral endopeptidase having an approximate molecular weight of 110000; (b) three acidic endopeptidases, with pH optima between 2.5 and 3.5, showing molecular and functional properties almost identical with those of the three proteinases previously purified from solubilized erythrocyte membranes [Pontremoli et al. (1979) Biochem. J. 181, 559 -5681; (c) two dipeptidylaminopeptidases whose molecular weights are around 80000 and tentatively identified as dipeptidyl aminopeptidases I1 and 111, respectively, on the basis of their substrate specificities and pH optima; (d) presumably two aminopeptidases, having an approximate molecular weight of 80 000 and classified as an aminopeptidase with broad substrate specificity and an aminopeptidase B, respectively. No evidence for any carboxypeptidase activity was found in the cytosolic compartment of mature human erythrocytes.The occurrence of proteolytic enzymes in human erythrocytes was originally described in 1953 by Morrison and Neurath [l]. Since then, several conflicting reports on this subject have appeared [2-51, mostly dealing with proteinase activities associated with the erythrocyte membrane [6]. The present state of the relevant research seems to indicate that three distinct acidic proteinases [7,8] are endowed in the membrane of mature erythrocytes.On the contrary, only few data are available concerning proteolytic activities in the soluble fraction of mature erythrocytes. Cytosolic erythrocyte arylamidases were described by Makinen and Makinen in 1971 [9] and two such aminopeptidases, specific for N-terminal arginine or lysine residues, have been recently purified and characterized by the same authors [lo]. MoreAbbreviation. CI~ACOH, trichloroacetic acid. Enzymes. Aminopeptidase (microsomal) (EC 3.4.1 1.1); dipeptidylaminopeptidase I1 (EC 3.4.14.2); serine carboxypeptidase (EC 3.14.16.1).over, the presence of dipeptidylaminopeptidases, more specifically indicated as dipeptidylaminopeptidase 111, has been reported [ll]. Another soluble proteolytic system has been described, but in rabbit reticulocytes and not in mature erythrocytes. This enzyme, which is characterized by its energy dependence, has been postulated to be responsible for the degradation of structurally abnorma...

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Section: Discussionsupporting

confidence: 76%

Identification of Proteolytic Activities in the Cytosolic Compartment of Mature Human Erythrocytes

Pontremoli

Melloni

Salamino

et al. 1980

European Journal of Biochemistry

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“…The two chains have molecular masses of 25 kDa (heavy chain) and Ͻ10 kDa (light chain) (25). Unlike most other cysteine proteases, which are small monomeric proteins with molecular masses of 20 -30 kDa, DPPI exists as an oligomeric enzyme of about 200 kDa, as determined by gel filtration (4,6,26,27). There are differing reports concerning the subunit composition of the enzyme, ranging from dimers to hexamers.…”

mentioning

confidence: 99%

“…DPPI is capable of sequentially removing dipeptides from the amino termini of various peptides and protein substrates (3)(4)(5)(6)(7) and, along with other cysteine proteases, is thought to play an important role in intracellular protein degradation and turnover (4,8,9). This enzyme has other postulated functions, including involvement in cell growth (10), neuraminidase activation (11), and platelet factor XIII activation (12).…”

mentioning

confidence: 99%

Molecular Cloning, Chromosomal Localization, and Expression of Murine Dipeptidyl Peptidase I

Pham

Armstrong

Zimonjic

et al. 1997

Journal of Biological Chemistry

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Dipeptidyl peptidase I (DPPI) is a lysosomal cysteine protease that catalyzes the sequential removal of dipeptides from the amino termini of various protein substrates. We have isolated a cDNA coding for murine DPPI from mouse thymus and spleen cDNA libraries. The deduced amino acid sequence codes for a protein of 462 amino acid residues; comparison of this deduced sequence with that of rat and human DPPI revealed 90.1% and 77.8% identity, respectively. Using DPPI cDNA, we obtained two BAC (Bacterial Artificial Chromosome) clones that contained the murine DPPI locus. The DPPI gene consists of seven exons and 6 introns, and spans approximately 20 kilobases. Using fluorescence in situ chromosome hybridization, we localized murine DPPI to chromosome 7D3-E1.1. We determined that DPPI protein is widely distributed in mouse tissues, although its relative abundance varies from tissue to tissue. In contrast to previous reports, we show here that DPPI mRNA and protein levels and enzymatic activity are unchanged during in vitro T cell activation, implying that this enzyme is not rate-limiting for granzyme processing.

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“…It is a potent competitive inhibitor of arylamidases (Marks, Datta, and Lajtha, 1968) but does not appreciably inhibit cathepsin B (Barrett and Poole, 1969). Although its mode of action on those arylamidases which act on amino acidnaphthylamides has not been clarified, both puromycin and the puromycin aminonucleoside inhibit the lysosomal dipeptidyl arylamidase II (McDonald, Reilly, Zeitman, and Ellis, 1968) by virtue of their size and their cationic charge. The physiological function of the amino acid and the dipeptidyl arylamidases is not known, but there has been a suggestion that they may de-activate regulatory peptides such as bradykinin (Behal, Little, and Klein, 1969).…”

Section: Contents Of Lysosomesmentioning

confidence: 99%

Lysosomal enzymes and inflammation with particular reference to rheumatoid diseases

Chayen¹,

Bitensky²

1972

Plastic and Reconstructive Surgery

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Leucine Naphthylamide: an Appropriate Substrate for the Histochemical Detection of Cathepsins B and B′

Cited by 59 publications

References 12 publications

Identification of Proteolytic Activities in the Cytosolic Compartment of Mature Human Erythrocytes

Identification of Proteolytic Activities in the Cytosolic Compartment of Mature Human Erythrocytes

Molecular Cloning, Chromosomal Localization, and Expression of Murine Dipeptidyl Peptidase I

Lysosomal enzymes and inflammation with particular reference to rheumatoid diseases

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