Purification of tyrosylprotein sulfotransferase from rat submandibular salivary glands

Samuel William, Patalapati Ramaprasad, Chinnaswam Kasinathan

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Tyrosylprotein sulfotransferase (TPST), the enzyme responsible for the sulfation of tyrosine residues, has been identified and characterized in submandibular salivary glands. In the present study, this enzyme was purified from the Golgi membranes of rat submandibular salivary glands using a Cibacron blue F3GA affinity column chromatography. Antibodies raised in rabbit against TPST detected the purified enzyme (50-54 kDa) and proteins consisting of molecular mass 50-54 kDa in the Golgi membranes of liver, submandibular salivary glands, stomach, cerebellum, thalamus, and pituitary. The protein levels in liver and salivary glands were higher compared to those found in the stomach, cerebellum, thalamus, and pituitary. The levels of immunoreactivity in cytosol and endoplasmic reticulum fractions of salivary glands were either undetectable or very low. The antibody was also used to immunoprecipitate the TPST activity and to isolate protein by immunoaffinity column. MnCl2 was required for the purified TPST. The enzyme exhibited optimum activity between pH 6.2 and 6.8 at 20 mM MnCl2. The apparent K(m) values of the purified enzyme for poly-(Glu6, Ala3, Tyr1) (EAY: M(r) 47,000) and 3'-phosphoadenosine 5'-phosphosulfate were 3 and 20 μM, respectively. The results presented here collectively demonstrate the purification of TPST and, for the first time, development of polyclonal antibody that recognizes this enzyme.

Original languageEnglish (US)
Pages (from-to)90-96
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume338
Issue number1
DOIs
StatePublished - Feb 1 1997

Fingerprint

Submandibular Gland
Salivary Glands
Purification
Rats
Enzymes
Cibacron Blue F 3GA
Thalamus
Liver
Cerebellum
Antibodies
Stomach
Phosphoadenosine Phosphosulfate
Membranes
Affinity chromatography
Column chromatography
Proteins
Molecular mass
Affinity Chromatography
Endoplasmic Reticulum
Cytosol

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

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title = "Purification of tyrosylprotein sulfotransferase from rat submandibular salivary glands",
abstract = "Tyrosylprotein sulfotransferase (TPST), the enzyme responsible for the sulfation of tyrosine residues, has been identified and characterized in submandibular salivary glands. In the present study, this enzyme was purified from the Golgi membranes of rat submandibular salivary glands using a Cibacron blue F3GA affinity column chromatography. Antibodies raised in rabbit against TPST detected the purified enzyme (50-54 kDa) and proteins consisting of molecular mass 50-54 kDa in the Golgi membranes of liver, submandibular salivary glands, stomach, cerebellum, thalamus, and pituitary. The protein levels in liver and salivary glands were higher compared to those found in the stomach, cerebellum, thalamus, and pituitary. The levels of immunoreactivity in cytosol and endoplasmic reticulum fractions of salivary glands were either undetectable or very low. The antibody was also used to immunoprecipitate the TPST activity and to isolate protein by immunoaffinity column. MnCl2 was required for the purified TPST. The enzyme exhibited optimum activity between pH 6.2 and 6.8 at 20 mM MnCl2. The apparent K(m) values of the purified enzyme for poly-(Glu6, Ala3, Tyr1) (EAY: M(r) 47,000) and 3'-phosphoadenosine 5'-phosphosulfate were 3 and 20 μM, respectively. The results presented here collectively demonstrate the purification of TPST and, for the first time, development of polyclonal antibody that recognizes this enzyme.",
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Purification of tyrosylprotein sulfotransferase from rat submandibular salivary glands. / William, Samuel; Ramaprasad, Patalapati; Kasinathan, Chinnaswam.

In: Archives of Biochemistry and Biophysics, Vol. 338, No. 1, 01.02.1997, p. 90-96.

Research output: Contribution to journalArticle

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