Purification and Characterization of a New Human Prostatic Acid Phosphatase Isoenzyme

Ming Fong Lin, Ching li Lee, Steven S.L. Li, T. Ming Chu

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

A new enzyme has been purified to homogeneity from human seminal plasma as determined by Polyacrylamide gel electrophoresis. This enzyme is shown to be an acid phosphatase (EC 3.1.3.2) by hydrolyzing a variety of phosphomonoesters at acidic pH, and hence designated as prostatic acid phosphatase II (PAP-II) to be differentiated from the conventional prostatic acid phosphatase (PAP) designated as PAP-I, which is isolated from the same source. PAP-II has a molecular weight (Mr) of 120 000 as estimated by gel filtration and possesses two subunits of Mr 55 000 each as revealed by sodium dodecyl sulfate-acrylamide gel electrophoresis, in comparison with molecular weights of 100 000 and 50 000, respectively, for PAP-I. The purified PAP-II demonstrates multiple pIs at 4.70–4.90, while those of PAP-I are at 4.84–5.33. PAP-II is inhibited by Fe3+, Ca2+, and La3+, whereas PAP-I is not affected at all. In addition to seminal plasma, PAP-II is detected only in tissue extracts of carcinoma prostate, benign prostate hypertrophy and normal prostate. Upon Immunoelectrophoresis, PAP-II shows a narrower precipitin arc with goat anti-PAP-I antiserum in reference to PAP-I, although an identical reactivity is detected by immunodiffusion. An immunological identity between PAP-I and PAP-II also is shown by their reactions with rabbit anti-PAP-II antiserum in immunodiffusion. Apart from these immunologic characteristics, results obtained from thermostability experiments, carbohydrate determination, in vivo clearance rate study, amino acid composition analysis, and peptide mapping data indicate that PAP-II is different from PAP-I. The vast difference in amino acid compositions rules out the possibilities that PAP-II and PAP-I are merely two distinctly different classes of glycosylated molecules and that PAP-II and PAP-I are the products of the same gene. It is concluded that PAP-II represents a new human PAP isoenzyme.

Original languageEnglish (US)
Pages (from-to)1055-1062
Number of pages8
JournalBiochemistry
Volume22
Issue number5
DOIs
StatePublished - Jan 1 1983

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Isoenzymes
Purification
prostatic acid phosphatase
Prostate
Immunodiffusion
Semen
Electrophoresis
Immune Sera
Molecular Weight
Gels
Molecular weight
Plasma (human)
Amino Acids
Precipitins
Immunoelectrophoresis
Peptide Mapping
Tissue Extracts
Acrylamide

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and Characterization of a New Human Prostatic Acid Phosphatase Isoenzyme. / Lin, Ming Fong; Lee, Ching li; Li, Steven S.L.; Chu, T. Ming.

In: Biochemistry, Vol. 22, No. 5, 01.01.1983, p. 1055-1062.

Research output: Contribution to journalArticle

Lin, Ming Fong ; Lee, Ching li ; Li, Steven S.L. ; Chu, T. Ming. / Purification and Characterization of a New Human Prostatic Acid Phosphatase Isoenzyme. In: Biochemistry. 1983 ; Vol. 22, No. 5. pp. 1055-1062.
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abstract = "A new enzyme has been purified to homogeneity from human seminal plasma as determined by Polyacrylamide gel electrophoresis. This enzyme is shown to be an acid phosphatase (EC 3.1.3.2) by hydrolyzing a variety of phosphomonoesters at acidic pH, and hence designated as prostatic acid phosphatase II (PAP-II) to be differentiated from the conventional prostatic acid phosphatase (PAP) designated as PAP-I, which is isolated from the same source. PAP-II has a molecular weight (Mr) of 120 000 as estimated by gel filtration and possesses two subunits of Mr 55 000 each as revealed by sodium dodecyl sulfate-acrylamide gel electrophoresis, in comparison with molecular weights of 100 000 and 50 000, respectively, for PAP-I. The purified PAP-II demonstrates multiple pIs at 4.70–4.90, while those of PAP-I are at 4.84–5.33. PAP-II is inhibited by Fe3+, Ca2+, and La3+, whereas PAP-I is not affected at all. In addition to seminal plasma, PAP-II is detected only in tissue extracts of carcinoma prostate, benign prostate hypertrophy and normal prostate. Upon Immunoelectrophoresis, PAP-II shows a narrower precipitin arc with goat anti-PAP-I antiserum in reference to PAP-I, although an identical reactivity is detected by immunodiffusion. An immunological identity between PAP-I and PAP-II also is shown by their reactions with rabbit anti-PAP-II antiserum in immunodiffusion. Apart from these immunologic characteristics, results obtained from thermostability experiments, carbohydrate determination, in vivo clearance rate study, amino acid composition analysis, and peptide mapping data indicate that PAP-II is different from PAP-I. The vast difference in amino acid compositions rules out the possibilities that PAP-II and PAP-I are merely two distinctly different classes of glycosylated molecules and that PAP-II and PAP-I are the products of the same gene. It is concluded that PAP-II represents a new human PAP isoenzyme.",
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N2 - A new enzyme has been purified to homogeneity from human seminal plasma as determined by Polyacrylamide gel electrophoresis. This enzyme is shown to be an acid phosphatase (EC 3.1.3.2) by hydrolyzing a variety of phosphomonoesters at acidic pH, and hence designated as prostatic acid phosphatase II (PAP-II) to be differentiated from the conventional prostatic acid phosphatase (PAP) designated as PAP-I, which is isolated from the same source. PAP-II has a molecular weight (Mr) of 120 000 as estimated by gel filtration and possesses two subunits of Mr 55 000 each as revealed by sodium dodecyl sulfate-acrylamide gel electrophoresis, in comparison with molecular weights of 100 000 and 50 000, respectively, for PAP-I. The purified PAP-II demonstrates multiple pIs at 4.70–4.90, while those of PAP-I are at 4.84–5.33. PAP-II is inhibited by Fe3+, Ca2+, and La3+, whereas PAP-I is not affected at all. In addition to seminal plasma, PAP-II is detected only in tissue extracts of carcinoma prostate, benign prostate hypertrophy and normal prostate. Upon Immunoelectrophoresis, PAP-II shows a narrower precipitin arc with goat anti-PAP-I antiserum in reference to PAP-I, although an identical reactivity is detected by immunodiffusion. An immunological identity between PAP-I and PAP-II also is shown by their reactions with rabbit anti-PAP-II antiserum in immunodiffusion. Apart from these immunologic characteristics, results obtained from thermostability experiments, carbohydrate determination, in vivo clearance rate study, amino acid composition analysis, and peptide mapping data indicate that PAP-II is different from PAP-I. The vast difference in amino acid compositions rules out the possibilities that PAP-II and PAP-I are merely two distinctly different classes of glycosylated molecules and that PAP-II and PAP-I are the products of the same gene. It is concluded that PAP-II represents a new human PAP isoenzyme.

AB - A new enzyme has been purified to homogeneity from human seminal plasma as determined by Polyacrylamide gel electrophoresis. This enzyme is shown to be an acid phosphatase (EC 3.1.3.2) by hydrolyzing a variety of phosphomonoesters at acidic pH, and hence designated as prostatic acid phosphatase II (PAP-II) to be differentiated from the conventional prostatic acid phosphatase (PAP) designated as PAP-I, which is isolated from the same source. PAP-II has a molecular weight (Mr) of 120 000 as estimated by gel filtration and possesses two subunits of Mr 55 000 each as revealed by sodium dodecyl sulfate-acrylamide gel electrophoresis, in comparison with molecular weights of 100 000 and 50 000, respectively, for PAP-I. The purified PAP-II demonstrates multiple pIs at 4.70–4.90, while those of PAP-I are at 4.84–5.33. PAP-II is inhibited by Fe3+, Ca2+, and La3+, whereas PAP-I is not affected at all. In addition to seminal plasma, PAP-II is detected only in tissue extracts of carcinoma prostate, benign prostate hypertrophy and normal prostate. Upon Immunoelectrophoresis, PAP-II shows a narrower precipitin arc with goat anti-PAP-I antiserum in reference to PAP-I, although an identical reactivity is detected by immunodiffusion. An immunological identity between PAP-I and PAP-II also is shown by their reactions with rabbit anti-PAP-II antiserum in immunodiffusion. Apart from these immunologic characteristics, results obtained from thermostability experiments, carbohydrate determination, in vivo clearance rate study, amino acid composition analysis, and peptide mapping data indicate that PAP-II is different from PAP-I. The vast difference in amino acid compositions rules out the possibilities that PAP-II and PAP-I are merely two distinctly different classes of glycosylated molecules and that PAP-II and PAP-I are the products of the same gene. It is concluded that PAP-II represents a new human PAP isoenzyme.

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