An agarose mercurial column for the separation of mercaptopapain and nonmercaptopapain

Cited by 203 publications

(69 citation statements)

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“…Papain (EC 3.4.22.2, from papaya latex, type III, Sigma) was repurified by covalent chromatography on an agarose mercurial column [9] leading to a 96070 active enzyme as determined by titration with E-64 [10].…”

Section: Methodsmentioning

confidence: 99%

Mechanism of inhibition of papain by chicken egg white cystatin

et al. 1989

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N-terminally truncated forms of chicken egg white cystatin and its cyanogen bromide fragments were isolated and assayed for inhibition of papain. Truncated forms beginning with Gly-9 and Ala-10 had a 5000-fold lower affinity for papain than the two isoelectric forms (pi=6.5 and 5.6) of the full-length inhibitor (K~--6 pM and 7 pM) or a truncated form beginning with Leu-7 (/~ = 6 pM), indicating the outstanding importance of one or two residues preceding conserved Gly-9 for binding. A weak inhibition of papain (K~ = 900 nM) was exhibited by the intermediate cyanogen bromide fragment (residues 30-89) containing the chicken cystatin QLVSG variation of the QWAG segment which is conserved in almost all members of the cystatin superfamily. The obtained affinity data provide independent evidence for the validity of the proposed docking model of a chicken cystatin-papain complex [( ) EMBO J. 7, 2593[( -2599.

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Section: Methodsmentioning

confidence: 99%

Mechanism of inhibition of papain by chicken egg white cystatin

et al. 1989

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“…Twice-crystallized papain was prepared by the method of Kimmel and Smith [S] from the protease (from Sigma) and further purified as described by Sluyterman and Wijdenes [9]. The purified enzyme (9.1 mg/ml) in a solution containing 10% dimethyl sulfoxide, 0.1 M KCl, 50 mM cysteine and 10 mM EDTA (pH adjusted to 7.0) was treated with a 5 molar excess of E-64-c (dissolved in a minimal amount of dimethyl sulfoxide).…”

Section: Methodsmentioning

confidence: 99%

Mode of binding of E‐64‐c, a potent thiol protease inhibitor, to papain as determined by X‐ray crystal analysis of the complex

et al. 1989

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The three‐dimensional structure of the E‐64‐c‐papain complex has been determined by X‐ray crystal analysis at 2.5 Å resolution (conventional R = 26.9%). The structure determined indicates that: (i) the C2 atom of the oxirane ring of E‐64‐c is covalently bound by the Sγ atom of Cys‐25 of papain; (ii) this covalent bond formation results in a configurational conversion of the oxirane C2 atom from the S‐ to the R‐form; and (iii) extensive hydrogen bonding and hydrophobic interactions are responsible for the specific interaction of the E‐64‐c molecule with papain.

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“…Papain (Worthington) was prepared by the method in [9]. L-Cysteine hydrochloride monohydrate (Sigma) was used without further purification.…”

Section: Methodsmentioning

confidence: 99%

Solvent isotope effects on tautomerization equilibria of papain and model thiolamines

Creighton

Schamp

1980

FEBS Letters

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An agarose mercurial column for the separation of mercaptopapain and nonmercaptopapain

Cited by 203 publications

References 9 publications

Mechanism of inhibition of papain by chicken egg white cystatin

Mechanism of inhibition of papain by chicken egg white cystatin

Mode of binding of E‐64‐c, a potent thiol protease inhibitor, to papain as determined by X‐ray crystal analysis of the complex

Solvent isotope effects on tautomerization equilibria of papain and model thiolamines

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