Pepsin Inhibitors from Ascaris lumbricoides

Abu-Erreish, Ghaleb M.; Peanasky, Robert J.

doi:10.1016/s0021-9258(19)42919-0

Cited by 46 publications

(6 citation statements)

References 27 publications

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“…Inhibitor released from the affinity gel had an average specific activity of 3070 units/mg of protein. This is a 750-fold increase in purification over the previous step and is comparable to the material of Abu-Erreish and Peanasky (1974a) after sequential column chromatography on Cellex-SE and DEAE-Sephadex. Affinity chromatography, unlike DEAE- Sephadex chromatography, did not separate the inhibitors into homogeneous fractions. This was undertaken by chromatofocusing (CF) chosen as a separation technique for its speed and reported high resolution and recoveries.…”

Section: Resultsmentioning

confidence: 56%

“…The overall recovery of pepsin inhibitors by the present affinity chromatography/chromatofocusing protocol (Table I) was 70% compared to 44% by the technique of Abu-Erreish and Peanasky (1974a). This increased yield was obtained by modifying the ultracentrifugation step and by replacing the sequential column chromatography steps on Bio-Gel P-30, Cellex-SE, and DEAE-cellulose with pepsin-AH-Sepharose affinity chromatography.…”

Section: Resultsmentioning

confidence: 81%

“…Cases of severe allergic reaction and anaphylactic shock as a result of exposure to A. suum have been reported (Stites et al, 1982) and must be considered in the design of purification protocols. The procedure of Abu-Erreish and Peanasky (1974a) for the initial preparation of pepsin inhibitors was followed with modifications. All steps were at 4 °C unless noted.…”

Section: Methodsmentioning

confidence: 99%

“…Visualized protein bands were positively identified as pepsin inhibitor by adaptation of the cytochemical staining technique of Furihata et al (1975). CF column fractions which contained more than one inhibitor species when analyzed by PAGE were further resolved by DEAE-Sephadex chromatography as previously described (Abu-Erreish & Peanasky, 1974a).…”

Section: Methodsmentioning

confidence: 99%

“…This potato protein does not show significant sequence homology with the inhibitor from Ascaris. The Ascaris inhibitor does not inhibit trypsin (Abu-Erreish & Peanasky, 1974a) or cathepsin D (Valler et al, 1985).…”

Section: Added In Proofmentioning

confidence: 95%

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Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum

Martzen¹,

McMullen²,

Smith³

et al. 1990

Biochemistry

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The major pepsin inhibitor from Ascaris suum was isolated by affinity chromatography and chromatofocusing. Its amino acid sequence was determined by automated Edman degradation of peptide fragments. Peptides were produced by chemical and enzymatic cleavage of pyridylethylated protein and were purified by reverse-phase high-performance liquid chromatography. The inhibitor consists of 149 residues with the following sequence: QFLFSMSTGP10FICTVKDNQV20FVANLPWTML30EGDDIQVGKE40 FAARVEDCTN50VKHDMAPTCT60KPPPFCGPQD70MKMFNFVGCS80VLGNKLFIDQ90KYVRDLTAK D100 HAEVQTFREK110IAAFEEQQEN120QPPSSGMPHG130AVPAGGLSPP140PPPSFCTVQ149. It has a molecular weight of 16,396. All cysteines are engaged as disulfide bonds: Cys(13)-Cys(59), Cys(48)-Cys(66), and Cys(79)-Cys(146). The protein is probably composed of two domains connected by a short hydrophobic region. This is the first aspartyl protease inhibitor of animal origin that has been sequenced. The sequence has no significant homology with any other known protein.

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Section: Resultsmentioning

confidence: 56%

Section: Resultsmentioning

confidence: 81%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Added In Proofmentioning

confidence: 95%

See 3 more Smart Citations

Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum

Martzen¹,

McMullen²,

Smith³

et al. 1990

Biochemistry

Self Cite

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Antigen processing for presentation by class II major histocompatibility complex requires cleavage by cathepsin E

et al. 1992

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Proteolytic degradation (processing) of antigen by antigen-presenting cells is a major regulatory step in the activation of a T lymphocyte immune response. However, the enzymes responsible for antigen processing remain largely undefined. In this study we show that cathepsin E, and not the ubiquitous lysosomal cathepsin D, is the major aspartic proteinase in a murine antigen-presenting cell line, A20. This enzyme is localized to a non-lysosomal compartment of the endosomal system in these cells. Functional studies using a highly specific inhibitor of cathepsin E show that this enzyme is essential for the processing of ovalbumin by this cell line. Thus, cathepsin E, whose function was hitherto unknown, may play a major role in antigen processing.

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Analysis of the interaction between the aspartic peptidase inhibitor SQAPI and aspartic peptidases using surface plasmon resonance

Farley

Christeller

Sullivan

et al. 2002

J of Molecular Recognition

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Aspartic peptidase inhibitors, which are themselves proteins, are strong inhibitors (small inhibition constants) of some aspartic peptidases but not others. However, there have been no studies of the kinetics of the interaction between a proteinaceous aspartic peptidase inhibitor and aspartic peptidases. This paper describes an analysis of rate constants for the interaction between recombinant squash aspartic peptidase inhibitor (rSQAPI) and a panel of aspartic peptidases that have a range of inhibition constants for SQAPI. Purified rSQAPI completely inhibits pepsin at a 1:1 molar ratio of pepsin to rSQAPI monomer (inhibition constant 1 nM). The interaction of pepsin with immobilized rSQAPI, at pH values between 3.0 and 6.0, was monitored using surface plasmon resonance. Binding of pepsin to rSQAPI was slow (association rate constants ca 10(4)M (-1)s(-1)), but rSQAPI was an effective pepsin inhibitor because dissociation of the rSQAPI-pepsin complex was much slower (dissociation rate constants ca 10(-4)s(-1)), especially at low pH values. Similar results were obtained with a His-tagged rSQAPI. Strong inhibition (inhibition constant 3 nM) of one isoform (rSap4) of the family of Candida albicans-secreted aspartic peptidases was, as with pepsin, characterized by slow binding of rSap4 and slower dissociation of the rSap4-inhibitor complex. In contrast, weaker inhibition of the Glomerella cingulata-secreted aspartic peptidase (inhibition constant 7 nM) and the C. albicans rSap1 and Sap2 isoenzymes (inhibition constants 25 and 400 nM, respectively) was, in each case, characterized by a larger dissociation rate constant.

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Pepsin Inhibitors from Ascaris lumbricoides

Cited by 46 publications

References 27 publications

Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum

Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum

Antigen processing for presentation by class II major histocompatibility complex requires cleavage by cathepsin E

Analysis of the interaction between the aspartic peptidase inhibitor SQAPI and aspartic peptidases using surface plasmon resonance

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