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Facile chemoenzymatic platform for protein tyrosine‐sulfate production.
Author(s) -
Lu Lu Yi,
Chen Bo Han,
Wu Jennifer YunShin,
Liu Tzu An,
Chen Da Huang,
Wang Chen Chu,
Lin Tsu Hsiang,
Hsu Ching Chen,
Yang Yuh Shyong
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.838.6
Subject(s) - sulfation , tyrosine , biochemistry , chemistry , enzyme , transmembrane protein , sulfotransferase , receptor
Protein tyrosine sulfation is a key post‐translational modification that mediates various critical functions, such as HIV entry, inflammation, coagulation, and sterility. Tyrosine O ‐sulfation is catalyzed by membrane‐associated tyrosylprotein sulfotransferase (TPST, EC 2.8.2.20), which is responsible for the sulfation of secreted and transmembrane proteins. The sulfated proteins are generally attained from tissue extraction, chemical synthesis, and enzymatic catalysis. Enzymatic catalysis provides direct, specific and easy method for the identification and preparation of sulfated proteins. Currently, TPST is mainly acquired from nature tissues or cell cultures. In this report, the constant source of active and homogeneous TPST at large quantity was achieved through prokaryotic expression. Furthermore, a continuous and cheap 3′‐phosphoadenosine‐5′‐phosphosulfate (PAPS) regenerating system was utilized to provide the activated sulfate for protein sulfation. This methodology rendered the increase in catalytic efficiency of TPST up to 10–100 folds than previous studies. Moreover, we applied this platform in vivo to produce target protein concurrently with sulfation on the specific tyrosine residue throughout bacterial cultivation. This platform will be beneficial to synthesize sulfated proteins and investigate the functions of protein sulfation. support: 98‐2627‐B‐009‐009