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N-linked glycosylation of CD38 is required for its structure stabilization but not for membrane localization

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Abstract

CD38 is a type II transmembrane protein with 25% of its molecular mass consisting of glycosyl moieties. It has long been predicted that the carbohydrate moieties of glycoproteins play important roles in the physical function and structural stability of the proteins on cell surfaces. To determine the structural/functional significance of glycosylation of the human CD38, the four potential N-linked glycosylation sites asparagine residues, N100, N164, N209 and N219 were mutated. The mutant (CD38mu) and wild-type (CD38wt) were expressed separately in Escherichia coli, HeLa, and MCF-7 cells. SDS-polyacrylamide gel electrophoresis under reducing conditions and western blotting indicated that the molecular mass of the CD38wt is 45 kDa, and that of the CD38mu is 34 kDa in HeLa cells. Importantly, the CD38mu protein expressed in HeLa cells, showed the high molecular weight oligomers in addition to the 34 kDa monomeric form. Similarly, in E. coli, the CD38wt formed dimers and other oligomers besides the monomeric form. Moreover, MCF-7 cells stably transfected with CD38wt cDNA, also revealed the presence of cross-linked oligomers when treated with a N-linked glycosylation inhibitor tunicamycin (TM). These results suggested that the N-linked glycosylation of CD38 plays a crucial role in the structure stability by preventing the formation inter-molecular cross-links. In addition, immunostaining, enzyme activity (cyclase), and western blotting data revealed that the glycosylation of human CD38 protein is not required for its localization to the cell membrane.

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Acknowledgments

This work was supported by a grant from the National Institutes Health, CA092115.

We thank Mr. Walter J. Pagel for his editorial review of the manuscript.

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Authors and Affiliations

  1. Department of Experimental Therapeutics, Unit 362, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA

    Yin Gao & Kapil Mehta

  2. Cancer Biology Program, Graduate School of Biomedical Sciences, The University of Texas, Houston, TX, USA

    Kapil Mehta

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  1. Yin Gao
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  2. Kapil Mehta
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Correspondence to Kapil Mehta.

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Gao, Y., Mehta, K. N-linked glycosylation of CD38 is required for its structure stabilization but not for membrane localization. Mol Cell Biochem 295, 1–7 (2007). https://doi.org/10.1007/s11010-006-9265-9

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  • DOI: https://doi.org/10.1007/s11010-006-9265-9

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