Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho

Jon D Wright; Sung-Wan An; Jian Xie; Joonho Yoon; Nicole Nischan; Jennifer J Kohler; Noelynn Oliver; Carmay Lim; Chou-Long Huang

doi:10.1096/fj.201700043R

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Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho

Journal article

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Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho

Jon D Wright, Sung-Wan An, Jian Xie, Joonho Yoon, Nicole Nischan, Jennifer J Kohler, Noelynn Oliver, Carmay Lim and Chou-Long Huang

The FASEB journal, Vol.31(8), pp.3574-3586

08/2017

DOI: 10.1096/fj.201700043R

PMCID: PMC5503716

PMID: 28442546

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Abstract

Soluble Klotho (sKlotho) is the shed ectodomain of antiaging membrane Klotho that contains 2 extracellular domains KL1 and KL2, each of which shares sequence homology to glycosyl hydrolases. sKlotho elicits pleiotropic cellular responses with a poorly understood mechanism of action. Notably, in injury settings, sKlotho confers cardiac and renal protection by down-regulating calcium-permeable transient receptor potential canonical type isoform 6 (TRPC6) channels in cardiomyocytes and glomerular podocytes. Inhibition of PI3K-dependent exocytosis of TRPC6 is thought to be the underlying mechanism, and recent studies showed that sKlotho interacts with α2-3-sialyllactose-containing gangliosides enriched in lipid rafts to inhibit raft-dependent PI3K signaling. However, the structural basis for binding and recognition of α2-3-sialyllactose by sKlotho is unknown. Using homology modeling followed by docking, we identified key protein residues in the KL1 domain that are likely involved in binding sialyllactose. Functional experiments based on the ability of Klotho to down-regulate TRPC6 channel activity confirm the importance of these residues. Furthermore, KL1 domain binds α2-3-sialyllactose, down-regulates TRPC6 channels, and exerts protection against stress-induced cardiac hypertrophy in mice. Our results support the notion that sialogangliosides and lipid rafts are membrane receptors for sKlotho and that the KL1 domain is sufficient for the tested biologic activities. These findings can help guide the design of a simpler Klotho mimetic.-Wright, J. D., An, S.-W., Xie, J., Yoon, J., Nischan, N., Kohler, J. J., Oliver, N., Lim, C., Huang, C.-L. Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho.

Membrane Microdomains

Humans

Lactose - analogs & derivatives

Glucuronidase - metabolism

Phosphatidylinositol 3-Kinases - metabolism

Sialic Acids - chemistry

Lactose - metabolism

Computer Simulation

Gangliosides - metabolism

HEK293 Cells

Binding Sites

Gangliosides - chemistry

Amino Acid Sequence

Sialic Acids - metabolism

Gene Expression Regulation - physiology

Models, Molecular

Phosphatidylinositol 3-Kinases - genetics

Animals

Glucuronidase - genetics

Lactose - chemistry

Protein Conformation

Signal Transduction - physiology

Mice

Carbohydrate Conformation

Cardiomegaly - metabolism

Details

Title: Subtitle: Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho
Creators: Jon D Wright - Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan
Sung-Wan An - Department of Internal Medicine, University of Texas Southwestern Medical Center, Texas, USA
Jian Xie - Department of Internal Medicine, University of Texas Southwestern Medical Center, Texas, USA
Joonho Yoon - Department of Internal Medicine, University of Texas Southwestern Medical Center, Texas, USA
Nicole Nischan - Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA
Jennifer J Kohler - Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA
Noelynn Oliver - Cardiometabolic Disease Research, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, USA
Carmay Lim - Department of Chemistry, National Tsing Hua University, HsinChu, Taiwan
Chou-Long Huang - Department of Internal Medicine, University of Texas Southwestern Medical Center, Texas, USA; chou-long.huang@utsouthwestern.edu
Resource Type: Journal article
Publication Details: The FASEB journal, Vol.31(8), pp.3574-3586
DOI: 10.1096/fj.201700043R
PMID: 28442546
PMCID: PMC5503716
NLM abbreviation: FASEB J
ISSN: 0892-6638
eISSN: 1530-6860
Grant note: R01 DK100605 / NIDDK NIH HHS R01 DK109887 / NIDDK NIH HHS
Language: English
Date published: 08/2017
Academic Unit: Nephrology; Internal Medicine
Record Identifier: 9984094382502771

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