Purification and characterization of dipeptidyl peptidase I from human spleen

McGuire, Michael J.; Lipsky, Peter E.; Thiele, Dwain L.

doi:10.1016/0003-9861(92)90519-3

Cited by 105 publications

(109 citation statements)

References 32 publications

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“…This also appears true for the human, where the level of expression of DPP-I transcripts observed in the current investigation is in close agreement with reports of the distribution of DPP-I enzyme activity in tissues (8) and hematopoietic cells (3,7). However, there are notable differences in the expression of DPP-I in the human and the rat.…”

Section: Identification Of Putative Regulatory Elements In the 5ј-flasupporting

confidence: 92%

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Human Dipeptidyl-peptidase I

Rao

Hoidal

1997

Journal of Biological Chemistry

146

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Dipeptidyl-peptidase I, a lysosomal cysteine proteinase, is important in intracellular degradation of proteins and appears to be a central coordinator for activation of many serine proteinases in immune/inflammatory cells. Little is known about the molecular genetics of the enzyme. In the present investigation the gene for dipeptidyl-peptidase I was cloned and characterized. The gene spans approximately 3.5 kilobases and consists of two exons and one intron. The genomic organization is distinct from the complex structures of the other members of the papain-type cysteine proteinase family. By fluorescence in situ hybridization, the gene was mapped to chromosomal region 11q14.1-q14.3. Analysis of the sequenced 5 -flanking region revealed no classical TATA or CCAAT box in the GC-rich region upstream of cap site. A number of possible regulatory elements that could account for tissue-specific expression were identified. Northern analyses demonstrated that the dipeptidyl-peptidase I message is expressed at high levels in lung, kidney, and placenta, at moderate to low levels in many organs, and at barely detectable levels in the brain, suggesting tissue-specific regulation. Among immune/inflammatory cells, the message is expressed at high levels in polymorphonuclear leukocytes and alveolar macrophages and their precursor cells. Treatment of lymphocytes with interleukin-2 resulted in a significant increase in dipeptidyl-peptidase I mRNA levels, suggesting that this gene is subjected to transcriptional regulation. The results provide initial insights into the molecular basis for the regulation of human dipeptidylpeptidase I.

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Section: Identification Of Putative Regulatory Elements In the 5ј-flasupporting

confidence: 92%

“…It is a lysosomal enzyme widely expressed in many tissues that is felt to be important in intracellular degradation of proteins. The enzyme was purified from human spleen and characterized as a glycoprotein with a pI of 5.4, a molecular mass of 200 kDa as determined by gel filtration under non-denaturing conditions, and a subunit size of 24 kDa (7).…”

mentioning

confidence: 99%

Human Dipeptidyl-peptidase I

Rao

Hoidal

1997

Journal of Biological Chemistry

146

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“…Cathepsin C is an oligomeric enzyme, each subunit consisting from three different polypeptide chains [4]. From 500 g of spleen we obtained 8 mg of cathepsin C. This is a much higher yield than those obtained by other methods, which included several chromatography steps [4,25,26]. The main advantage of this procekDa 93 - Fig.…”

Section: Resultsmentioning

confidence: 93%

Interaction of human cathepsin C with chicken cystatin

et al. 1996

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Cathepsin C was purified from human spleen by a rapid procedure, which included homogenization, ammonium sulfate precipitation, gel filtration on Sephacryl S-200 and fmally affinity chromatography on chicken cystatin-Sepharose. The interaction between cathepsin C and chicken cystatin was further characterized. It was found to be accompanied by a maximum decrease in fluorescence emission intensity at 330 nm. Fluorescence titration showed that human catbepsin C can bind four chicken cystatin molecules. The 4:1 binding stoichiometry was confirmed by titration monitored by the loss of enzyme activity. A non-competitive-competitive type of inhibition was determined from a double-reciprocal Lineweaver-Burk plot with a Kj value of 0.22 nM for the non-competitive inhibition.

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“…As demonstrated for elastase, cathepsin G and lymphocyte granzymes, the cleavage of a two-amino-acid residue propeptide is required for PR3 to assume its conformation dependent enzymatic function [29,30]. Its cleavage is performed by dipeptidylpeptidase I (DPPI) [31,32], which appears to be missing in nonhematopoetic eukaryotic cells [30,33]. In U937 cells the activation dipeptide of PR3 appears to be cleaved by a cysteine protease distinct from DPPI, while elastase is activated by DPPI [26].…”

Section: Enzymatic Activity Of Rpr3mentioning

confidence: 99%

Recombinant human proteinase 3, the Wegener's autoantigen, expressed in HMC‐1 cells is enzymatically active and recognized by c‐ANCA

Specks¹,

Fass²,

Fautsch³

et al. 1996

FEBS Letters

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We developed a stable expression system for conformationally intact recombinant human PR3 (rPR3) using the human mast cell line HMC-I. Like in U937 cells, the rPR3 is processed from a 34 kDa precursor to the 29 kDa mature form, primarily as the result of oligosaccharide trimming. The rPR3 binds [3H]DFP and hydrolyzes the snbstrate N-methoxysuccinylAla-Ala-Pro-Val-pNA. The enzymatic activity is inhibited by greater than 95% by al-PI. The rPR3 and the enzymatically inactive mutant rPR3-S176A are both packaged in granules. Thus, proteolytic autoprocessing is not required for PRYs targeting to granules. This rPR3 is the first to be recognized by most c-ANCA from WG patients and all anti-PR3 ANCA that were detected by standard anti-PR3 specific ELISA. This expression system for rPR3 represents a versatile tool for the analysis of its intraceHular processing, structure-function relationships and interaction with autoantibodies.

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Purification and characterization of dipeptidyl peptidase I from human spleen

Cited by 105 publications

References 32 publications

Human Dipeptidyl-peptidase I

Human Dipeptidyl-peptidase I

Interaction of human cathepsin C with chicken cystatin

Recombinant human proteinase 3, the Wegener's autoantigen, expressed in HMC‐1 cells is enzymatically active and recognized by c‐ANCA

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