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An LPL-specific monoclonal antibody, 88B8, that abolishes the binding of LPL to GPIHBP1.

ABSTRACT: LPL contains two principal domains: an amino-terminal catalytic domain (residues 1-297) and a carboxyl-terminal domain (residues 298-448) that is important for binding lipids and binding glycosylphosphatidylinositol-anchored high density lipoprotein binding protein 1 (GPIHBP1) (an endothelial cell protein that shuttles LPL to the capillary lumen). The LPL sequences required for GPIHBP1 binding have not been examined in detail, but one study suggested that sequences near LPL's carboxyl terminus (residues ?403-438) were crucial. Here, we tested the ability of LPL-specific monoclonal antibodies (mAbs) to block the binding of LPL to GPIHBP1. One antibody, 88B8, abolished LPL binding to GPIHBP1. Consistent with those results, antibody 88B8 could not bind to GPIHBP1-bound LPL on cultured cells. Antibody 88B8 bound poorly to LPL proteins with amino acid substitutions that interfered with GPIHBP1 binding (e.g., C418Y, E421K). However, the sequences near LPL's carboxyl terminus (residues ?403-438) were not sufficient for 88B8 binding; upstream sequences (residues 298-400) were also required. Additional studies showed that these same sequences are required for LPL binding to GPIHBP1. In conclusion, we identified an LPL mAb that binds to LPL's GPIHBP1-binding domain. The binding of both antibody 88B8 and GPIHBP1 to LPL depends on large segments of LPL's carboxyl-terminal domain.

SUBMITTER: Allan CM

PROVIDER: S-EPMC5036369 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

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An LPL-specific monoclonal antibody, 88B8, that abolishes the binding of LPL to GPIHBP1.

Allan Christopher M CM Larsson Mikael M Hu Xuchen X He Cuiwen C Jung Rachel S RS Mapar Alaleh A Voss Constance C Miyashita Kazuya K Machida Tetsuo T Murakami Masami M Nakajima Katsuyuki K Bensadoun André A Ploug Michael M Fong Loren G LG Young Stephen G SG Beigneux Anne P AP

Journal of lipid research 20160805 10

LPL contains two principal domains: an amino-terminal catalytic domain (residues 1-297) and a carboxyl-terminal domain (residues 298-448) that is important for binding lipids and binding glycosylphosphatidylinositol-anchored high density lipoprotein binding protein 1 (GPIHBP1) (an endothelial cell protein that shuttles LPL to the capillary lumen). The LPL sequences required for GPIHBP1 binding have not been examined in detail, but one study suggested that sequences near LPL's carboxyl terminus ( ...[more]

PMID: 27494936

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Dataset Information

An LPL-specific monoclonal antibody, 88B8, that abolishes the binding of LPL to GPIHBP1.

Publications

An LPL-specific monoclonal antibody, 88B8, that abolishes the binding of LPL to GPIHBP1.

Similar Datasets

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