Hieronymain I, a new cysteine peptidase isolated from unripe fruits of Bromelia hieronymi Mez (Bromeliaceae)

A new peptidase, named hieronymain I, was purified to homogeneity from unripe fruits of Bromelia hieronymi Mez (Bromeliaceae) by acetone fractionation followed by cation exchange chromatography (FPLC) on CM-Sepharose FF. Homogeneity of the enzyme was confirmed by mass spectroscopy (MALDI-TOF), isoel...

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Bibliographic Details
Published inJournal of Protein Chemistry Vol. 22; no. 2; pp. 127 - 134
Main Authors Bruno, Mariela A, Pardo, Marcelo F, Caffini, Néstor O, López, Laura M I
Format Journal Article
LanguageEnglish
Published United States Springer Nature B.V 01.02.2003
Subjects
Amino Acid Sequence
Bromelia
Bromeliaceae
Bromeliaceae - enzymology
Casein
Cation exchange
Cation exchanging
Chromatography
Chromatography, Ion Exchange
Cysteine
Cysteine Endopeptidases - chemistry
Cysteine Endopeptidases - isolation & purification
Cysteine Endopeptidases - metabolism
Cysteine Proteinase Inhibitors - pharmacology
Electrophoresis, Polyacrylamide Gel
Enzyme Stability
Enzymes
Fractionation
Fruit - chemistry
Fruit - enzymology
Fruits
Homogeneity
Iodoacetic acid
Isoelectric Focusing
Liquid chromatography
Mass Spectrometry
Mass spectroscopy
Molecular Sequence Data
Molecular Weight
Peptidase
Peptidases
Peptides
Plant Extracts - chemistry
Proteases
Proteins
Proteolysis
Sequence Homology, Amino Acid
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Summary:A new peptidase, named hieronymain I, was purified to homogeneity from unripe fruits of Bromelia hieronymi Mez (Bromeliaceae) by acetone fractionation followed by cation exchange chromatography (FPLC) on CM-Sepharose FF. Homogeneity of the enzyme was confirmed by mass spectroscopy (MALDI-TOF), isoelectric focusing, and SDS-PAGE. Hieronymain is a basic peptidase (pI > 9.3) and its molecular mass was 24,066 Da. Maximum proteolytic activity on casein (>90% of maximum activity) was achieved at pH 8.5-9.5. The enzyme was completely inhibited by E-64 and iodoacetic acid and activated by the addition of cysteine; these results strongly suggest that the isolated protease should be included within the cysteine group. The N-terminal sequence of hieronymain (ALPESIDWRAKGAVTEVKRQDG) was compared with 25 plant cysteine proteases that showed more than 50% of identity.
Bibliography:ObjectType-Article-2
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ISSN:0277-8033
1572-3887
1573-4943
DOI:10.1023/A:1023418812832