Conformation-Sensing Antibodies Stabilize the Oxidized Form of PTP1B and Inhibit Its Phosphatase Activity

被引：124

作者：

Haque, Aftabul ^{[1
,2
]}

Andersen, Jannik N. ^{[1
]}

Salmeen, Annette ^{[3
]}

Barford, David ^{[3
]}

Tonks, Nicholas K. ^{[1
]}

机构：

[1] Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA

[2] SUNY Stony Brook, Mol & Cellular Biol Grad Program, Stony Brook, NY 11790 USA

[3] Inst Canc Res, Chester Beatty Labs, Div Struct Biol, London SW3 6JB, England

来源：

CELL | 2011年 / 147卷 / 01期

关键词：

PROTEIN-TYROSINE-PHOSPHATASE; SIGNAL-TRANSDUCTION; INSULIN-RECEPTOR; IN-VIVO; REVERSIBLE OXIDATION; HYDROGEN-PEROXIDE; REDOX REGULATION; 1B; DEPHOSPHORYLATION; MICE;

D O I：

10.1016/j.cell.2011.08.036

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Protein tyrosine phosphatase 1B (PTP1B) plays important roles in downregulation of insulin and leptin signaling and is an established therapeutic target for diabetes and obesity. PTP1B is regulated by reactive oxygen species (ROS) produced in response to various stimuli, including insulin. The reversibly oxidized form of the enzyme (PTP1B-OX) is inactive and undergoes profound conformational changes at the active site. We generated conformation-sensor antibodies, in the form of single-chain variable fragments (scFvs), that stabilize PTP1B-OX and thereby inhibit its phosphatase function. Expression of conformation-sensor scFvs as intracellular antibodies (intrabodies) enhanced insulin-induced tyrosyl phosphorylation of the beta subunit of the insulin receptor and its substrate IRS-1 and increased insulin-induced phosphorylation of PKB/AKT. Our data suggest that stabilization of the oxidized, inactive form of PTP1B with appropriate therapeutic molecules may offer a paradigm for phosphatase drug development.

引用

页码：185 / 198

页数：14

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