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Comparative Mechanistic and Substrate Specificity Study of Inositol Polyphosphate 5-Phosphatase Schizosaccharomyces pombe Synaptojanin and SHIP2
Journal article   Open access   Peer reviewed

Comparative Mechanistic and Substrate Specificity Study of Inositol Polyphosphate 5-Phosphatase Schizosaccharomyces pombe Synaptojanin and SHIP2

Yuling Chi, Bo Zhou, Wei-Qing Wang, Sung-Kee Chung, Yong-Uk Kwon, Young-Hoon Ahn, Young-Tae Chang, Yosuke Tsujishita, James Hurley and Zhong-Yin Zhang
The Journal of biological chemistry, v 279(43), pp 44987-44995
22 Oct 2004
PMID: 15316017
url
http://www.jbc.org/content/279/43/44987.full.pdfView
Published, Version of Record (VoR) Open
url
https://doi.org/10.1074/jbc.M406416200View
Published, Version of Record (VoR) Open

Abstract

Schizosaccharomyces pombe
Inositol-5-phosphatases are important enzymes involved in the regulation of diverse cellular processes from synaptic vesicle recycling to insulin signaling. We describe a comparative study of two representative inositol-5-phosphatases, Schizosaccharomyces pombe synaptojanin (SPsynaptojanin) and human SH2 domain- containing inositol-5-phosphatase SHIP2. We show that in addition to Mg super(2+), transition metals such as Mn super(2+), Co super(2+), and Ni super(2+) are also effective activators of SPsynaptojanin. In contrast, Ca super(2+) and Cu super(2+) are inhibitory. We provide evidence that Mg super(2+) binds the same site occupied by Ca super(2+) observed in the crystal structure of SPsynaptojanin complexed with inositol 1,4- bisphosphate (Ins(1,4)P sub(2)). Ionizations important for substrate binding and catalysis are defined for the SPsynaptojanin-catalyzed Ins(1,4,5)P sub(3) reaction. Kinetic analysis with four phosphatidylinositol lipids bearing a 5- phosphate and 54 water-soluble inositol phosphates reveals that SP-synaptojanin and SHIP2 possess much broader substrate specificity than previously appreciated. The rank order for SPsynaptojanin is Ins(2,4,5)P sub(3) > phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P sub(2)) ~ Ins(4,5)P sub(2) ~ Ins(1,4,5)P sub(3) ~ Ins(4,5,6)P sub(3) > PtdIns(3,5)P sub(2) ~ PtdIns(3,4,5)P sub(3) ~ Ins(1,2,4,5)P sub(4) ~ Ins(1,3,4,5)P sub(4) ~ Ins-(2,4,5,6)P sub(4) ~ Ins(1,2,4,5,6)P sub(5). The rank order for SHIP2 is Ins(1,2,3,4,5)P sub(5) > Ins(1,3,4,5)P sub(4) > PtdIns(3,4,5)P sub(4) ~ PtdIns(3,5)P sub(2) ~ Ins(1,4,5,6)P sub(4) ~ Ins(2,4,5,6)P sub(4). Because inositol phosphate isomers elicit different biological activities, the extended substrate specificity for SPsynaptojanin and SHIP2 suggest that these enzymes likely have multiple roles in cell signaling and may regulate distinct pathways. The unique substrate specificity profiles and the importance of 2-position phosphate in binding also have important implications for the design of potent and selective SPsynaptojanin and SHIP2 inhibitors for pharmacological investigation.

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Biochemistry & Molecular Biology
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