Enhanced selectivity for sulfatide by engineered human glycolipid transfer protein

Valeria R. Samygina; Alexander N. Popov; Aintzane Cabo-Bilbao; Borja Ochoa-Lizarralde; Felipe Goni-De-Cerio; Xiuhong Zhai; Julian G. Molotkovsky; Dinshaw J. Patel; Rhoderick E. Brown; Lucy Malinina

doi:10.1016/j.str.2011.09.011

Enhanced selectivity for sulfatide by engineered human glycolipid transfer protein

Valeria R. Samygina, Alexander N. Popov, Aintzane Cabo-Bilbao, Borja Ochoa-Lizarralde, Felipe Goni-De-Cerio, Xiuhong Zhai, Julian G. Molotkovsky, Dinshaw J. Patel, Rhoderick E. Brown, Lucy Malinina

The Hormel Institute

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

Human glycolipid transfer protein (GLTP) fold represents a novel structural motif for lipid binding/transfer and reversible membrane translocation. GLTPs transfer glycosphingolipids (GSLs) that are key regulators of cell growth, division, surface adhesion, and neurodevelopment. Herein, we report structure-guided engineering of the lipid binding features of GLTP. New crystal structures of wild-type GLTP and two mutants (D48V and A47DD48V), each containing bound N-nervonoyl-sulfatide, reveal the molecular basis for selective anchoring of sulfatide (3-O-sulfo-galactosylceramide) by D48V-GLTP. Directed point mutations of "portal entrance" residues, A47 and D48, reversibly regulate sphingosine access to the hydrophobic pocket via a mechanism that could involve homodimerization. "Door-opening" conformational changes by phenylalanines within the hydrophobic pocket are revealed during lipid encapsulation by new crystal structures of bona fide apo-GLTP and GLTP complexed with N-oleoyl-glucosylceramide. The development of "engineered GLTPs" with enhanced specificity for select GSLs provides a potential new therapeutic approach for targeting GSL-mediated pathologies.

Original language	English (US)
Pages (from-to)	1644-1654
Number of pages	11
Journal	Structure
Volume	19
Issue number	11
DOIs	https://doi.org/10.1016/j.str.2011.09.011
State	Published - Nov 9 2011

Bibliographical note

Funding Information:
This research was supported by Spanish Ministerio de Ciencia e Innovacion (MICINN BFU2010-17711), Russian Foundation for Basic Research (#09-04-00313), USPHS (NIGMS GM45928 and NCI CA121493), the Abby Rockefeller Mauzé Trust, and Maloris and Hormel Foundations. The authors thank Margarita Malakhova for helpful advice, the personnel at ESRF beamline ID 23-1 and ID 14-4 for assistance with X-ray data collection, our colleagues Sandra Delgado, Jevgenia Tamjar, Oscar Millet, and Inma Gómez García from CICbioGUNE for help with fluorescence and DLS measurements, and Helen Pike from the UMN-Hormel Institute for expressing and purifying protein used for transfer activity measurements.

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title = "Enhanced selectivity for sulfatide by engineered human glycolipid transfer protein",

abstract = "Human glycolipid transfer protein (GLTP) fold represents a novel structural motif for lipid binding/transfer and reversible membrane translocation. GLTPs transfer glycosphingolipids (GSLs) that are key regulators of cell growth, division, surface adhesion, and neurodevelopment. Herein, we report structure-guided engineering of the lipid binding features of GLTP. New crystal structures of wild-type GLTP and two mutants (D48V and A47DD48V), each containing bound N-nervonoyl-sulfatide, reveal the molecular basis for selective anchoring of sulfatide (3-O-sulfo-galactosylceramide) by D48V-GLTP. Directed point mutations of {"}portal entrance{"} residues, A47 and D48, reversibly regulate sphingosine access to the hydrophobic pocket via a mechanism that could involve homodimerization. {"}Door-opening{"} conformational changes by phenylalanines within the hydrophobic pocket are revealed during lipid encapsulation by new crystal structures of bona fide apo-GLTP and GLTP complexed with N-oleoyl-glucosylceramide. The development of {"}engineered GLTPs{"} with enhanced specificity for select GSLs provides a potential new therapeutic approach for targeting GSL-mediated pathologies.",

author = "Samygina, {Valeria R.} and Popov, {Alexander N.} and Aintzane Cabo-Bilbao and Borja Ochoa-Lizarralde and Felipe Goni-De-Cerio and Xiuhong Zhai and Molotkovsky, {Julian G.} and Patel, {Dinshaw J.} and Brown, {Rhoderick E.} and Lucy Malinina",

note = "Funding Information: This research was supported by Spanish Ministerio de Ciencia e Innovacion (MICINN BFU2010-17711), Russian Foundation for Basic Research (#09-04-00313), USPHS (NIGMS GM45928 and NCI CA121493), the Abby Rockefeller Mauz{\'e} Trust, and Maloris and Hormel Foundations. The authors thank Margarita Malakhova for helpful advice, the personnel at ESRF beamline ID 23-1 and ID 14-4 for assistance with X-ray data collection, our colleagues Sandra Delgado, Jevgenia Tamjar, Oscar Millet, and Inma G{\'o}mez Garc{\'i}a from CICbioGUNE for help with fluorescence and DLS measurements, and Helen Pike from the UMN-Hormel Institute for expressing and purifying protein used for transfer activity measurements. ",

year = "2011",

month = nov,

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doi = "10.1016/j.str.2011.09.011",

language = "English (US)",

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T1 - Enhanced selectivity for sulfatide by engineered human glycolipid transfer protein

AU - Samygina, Valeria R.

AU - Popov, Alexander N.

AU - Cabo-Bilbao, Aintzane

AU - Ochoa-Lizarralde, Borja

AU - Goni-De-Cerio, Felipe

AU - Zhai, Xiuhong

AU - Molotkovsky, Julian G.

AU - Patel, Dinshaw J.

AU - Brown, Rhoderick E.

AU - Malinina, Lucy

N1 - Funding Information: This research was supported by Spanish Ministerio de Ciencia e Innovacion (MICINN BFU2010-17711), Russian Foundation for Basic Research (#09-04-00313), USPHS (NIGMS GM45928 and NCI CA121493), the Abby Rockefeller Mauzé Trust, and Maloris and Hormel Foundations. The authors thank Margarita Malakhova for helpful advice, the personnel at ESRF beamline ID 23-1 and ID 14-4 for assistance with X-ray data collection, our colleagues Sandra Delgado, Jevgenia Tamjar, Oscar Millet, and Inma Gómez García from CICbioGUNE for help with fluorescence and DLS measurements, and Helen Pike from the UMN-Hormel Institute for expressing and purifying protein used for transfer activity measurements.

PY - 2011/11/9

Y1 - 2011/11/9

N2 - Human glycolipid transfer protein (GLTP) fold represents a novel structural motif for lipid binding/transfer and reversible membrane translocation. GLTPs transfer glycosphingolipids (GSLs) that are key regulators of cell growth, division, surface adhesion, and neurodevelopment. Herein, we report structure-guided engineering of the lipid binding features of GLTP. New crystal structures of wild-type GLTP and two mutants (D48V and A47DD48V), each containing bound N-nervonoyl-sulfatide, reveal the molecular basis for selective anchoring of sulfatide (3-O-sulfo-galactosylceramide) by D48V-GLTP. Directed point mutations of "portal entrance" residues, A47 and D48, reversibly regulate sphingosine access to the hydrophobic pocket via a mechanism that could involve homodimerization. "Door-opening" conformational changes by phenylalanines within the hydrophobic pocket are revealed during lipid encapsulation by new crystal structures of bona fide apo-GLTP and GLTP complexed with N-oleoyl-glucosylceramide. The development of "engineered GLTPs" with enhanced specificity for select GSLs provides a potential new therapeutic approach for targeting GSL-mediated pathologies.

AB - Human glycolipid transfer protein (GLTP) fold represents a novel structural motif for lipid binding/transfer and reversible membrane translocation. GLTPs transfer glycosphingolipids (GSLs) that are key regulators of cell growth, division, surface adhesion, and neurodevelopment. Herein, we report structure-guided engineering of the lipid binding features of GLTP. New crystal structures of wild-type GLTP and two mutants (D48V and A47DD48V), each containing bound N-nervonoyl-sulfatide, reveal the molecular basis for selective anchoring of sulfatide (3-O-sulfo-galactosylceramide) by D48V-GLTP. Directed point mutations of "portal entrance" residues, A47 and D48, reversibly regulate sphingosine access to the hydrophobic pocket via a mechanism that could involve homodimerization. "Door-opening" conformational changes by phenylalanines within the hydrophobic pocket are revealed during lipid encapsulation by new crystal structures of bona fide apo-GLTP and GLTP complexed with N-oleoyl-glucosylceramide. The development of "engineered GLTPs" with enhanced specificity for select GSLs provides a potential new therapeutic approach for targeting GSL-mediated pathologies.

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JF - Structure

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Enhanced selectivity for sulfatide by engineered human glycolipid transfer protein

Abstract

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