Affinity, Kinetic, and Structural Study of the Interaction of 3-O-Sulfotransferase Isoform 1 with Heparan Sulfate
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- 作者：Eva Muñ ; oz ; Ding Xu ; Melissa Kemp ; Fuming Zhang ; Jian Liu ; and Robert J. Linhardt
- 刊名：Biochemistry
- 出版年：2006
- 出版时间：April 25, 2006
- 年：2006
- 卷：45
- 期：16
- 页码：5122 - 5128
- 全文大小：218K
- 年卷期：v.45,no.16(April 25, 2006)
- ISSN：1520-4995
文摘
The 3-O-sulfonation of glucosamine residues in heparan sulfate (HS) by 3-O-sulfotransferase(3-OST) is a key substitution that is present in HS sequences of biological importance, in particular HSanticoagulant activity. Six different isoforms of 3-OST have been identified that exhibit different substratespecificity. In this paper the affinity and kinetics of the interaction between 3-O-sulfotransferase isoform1 (3-OST-1) and HS have been examined using surface plasmon resonance (SPR). 3-OST-1 binds withmicomolar affinity to HS (KD = 2.79 
M), and this interaction is apparently independent of the presenceof the coenzyme, 3'-phosphoadenosine 5'-phosphosulfate (PAPS). A conformational change in the complexhas also been detected, supporting data from previous studies. Selected 3-OST-1 mutants have providedvaluable information of amino acid residues that participate in 3-OST-1 interaction with HS substrate andits catalytic activity. The results from this study contribute to understanding the substrate specificity amongthe 3-OST isoforms and in the mechanism of 3-OST-1-catalyzed biosynthesis of anticoagulant HS.
M), and this interaction is apparently independent of the presenceof the coenzyme, 3'-phosphoadenosine 5'-phosphosulfate (PAPS). A conformational change in the complexhas also been detected, supporting data from previous studies. Selected 3-OST-1 mutants have providedvaluable information of amino acid residues that participate in 3-OST-1 interaction with HS substrate andits catalytic activity. The results from this study contribute to understanding the substrate specificity amongthe 3-OST isoforms and in the mechanism of 3-OST-1-catalyzed biosynthesis of anticoagulant HS.