Cellular Localization and Biochemical Analysis of Mammalian CDC50A, a Glycosylated β-subunit for P4 ATPases

Overview

Journal J Histochem Cytochem

Publisher Sage Publications

Specialty Biochemistry

Date 2012 Jan 19

PMID 22253360

Citations 13

Authors

Dineke E Folmer

Kam S Mok

Sebastiaan W de Wee

Suzanne Duijst

Johan K Hiralall

Jurgen Seppen

Ronald P J Oude Elferink

Coen C Paulusma

Affiliations

Soon will be listed here.

Abstract

CDC50 proteins are β-subunits for P4 ATPases, which upon heterodimerization form a functional phospholipid translocation complex. Emerging evidence in mouse models and men links mutations in P4 ATPase genes with human disease. This study analyzed the tissue distribution and cellular localization of CDC50A, the most abundant and ubiquitously expressed CDC50 homologue in the mouse. The authors have raised antibodies that detect mouse and human CDC50A and studied CDC50A localization and glycosylation status in mouse liver cells. CDC50A is a terminal-glycosylated glycoprotein and is expressed in hepatocytes and liver sinusoidal endothelial cells, where it resides in detergent-resistant membranes. In pancreas and stomach, CDC50A localized to secretory vesicles, whereas in the kidney, CDC50A localized to the apical region of proximal convoluted tubules of the cortex. In WIF-B9 cells, CDC50A partially costains with the trans-Golgi network. Data suggest that CDC50A is present as a fully glycosylated protein in vivo, which presumes interaction with distinct P4 ATPases.

Citing Articles

Knockdown of TMEM30A in renal tubular epithelial cells leads to reduced glucose absorption.

Chen S, Song X, Xiao Q, Wang L, Zhu X, Zou Y BMC Nephrol. 2023; 24(1):250.

PMID: 37612668 PMC: 10464243. DOI: 10.1186/s12882-023-03299-8.

Physiological and Pathological Functions of TMEM30A: An Essential Subunit of P4-ATPase Phospholipid Flippases.

Li J, Zhao Y, Wang N J Lipids. 2023; 2023:4625567.

PMID: 37200892 PMC: 10188266. DOI: 10.1155/2023/4625567.

Loss of phosphatidylserine flippase β-subunit Tmem30a in podocytes leads to albuminuria and glomerulosclerosis.

Liu W, Peng L, Tian W, Li Y, Zhang P, Sun K Dis Model Mech. 2021; 14(6).

PMID: 34080006 PMC: 8246268. DOI: 10.1242/dmm.048777.

The phosphatidylserine flippase β-subunit Tmem30a is essential for normal insulin maturation and secretion.

Yang Y, Sun K, Liu W, Li X, Tian W, Shuai P Mol Ther. 2021; 29(9):2854-2872.

PMID: 33895325 PMC: 8417432. DOI: 10.1016/j.ymthe.2021.04.026.

Disruption of Tmem30a results in cerebellar ataxia and degeneration of Purkinje cells.

Yang Y, Sun K, Liu W, Zhang L, Peng K, Zhang S Cell Death Dis. 2018; 9(9):899.

PMID: 30185775 PMC: 6125289. DOI: 10.1038/s41419-018-0938-6.

References

Siggs O, Arnold C, Huber C, Pirie E, Xia Y, Lin P . The P4-type ATPase ATP11C is essential for B lymphopoiesis in adult bone marrow. Nat Immunol. 2011; 12(5):434-40. PMC: 3079768. DOI: 10.1038/ni.2012. View

Eppens E, van Mil S, de Vree J, Mok K, Juijn J, Oude Elferink R . FIC1, the protein affected in two forms of hereditary cholestasis, is localized in the cholangiocyte and the canalicular membrane of the hepatocyte. J Hepatol. 2001; 35(4):436-43. DOI: 10.1016/s0168-8278(01)00158-1. View

Koenderink J, Geibel S, Grabsch E, de Pont J, Bamberg E, Friedrich T . Electrophysiological analysis of the mutated Na,K-ATPase cation binding pocket. J Biol Chem. 2003; 278(51):51213-22. DOI: 10.1074/jbc.M306384200. View

Harder R, Uhlig H, Kashan A, Schutt B, Duijvestijn A, Butcher E . Dissection of murine lymphocyte-endothelial cell interaction mechanisms by SV-40-transformed mouse endothelial cell lines: novel mechanisms mediating basal binding, and alpha 4-integrin-dependent cytokine-induced adhesion. Exp Cell Res. 1991; 197(2):259-67. DOI: 10.1016/0014-4827(91)90431-s. View

Halleck M, Lawler JF J, Blackshaw S, Gao L, Nagarajan P, Hacker C . Differential expression of putative transbilayer amphipath transporters. Physiol Genomics. 2000; 1(3):139-50. DOI: 10.1152/physiolgenomics.1999.1.3.139. View

Yabas M, Teh C, Frankenreiter S, Lal D, Roots C, Whittle B . ATP11C is critical for the internalization of phosphatidylserine and differentiation of B lymphocytes. Nat Immunol. 2011; 12(5):441-9. PMC: 3272780. DOI: 10.1038/ni.2011. View

Folmer D, Oude Elferink R, Paulusma C . P4 ATPases - lipid flippases and their role in disease. Biochim Biophys Acta. 2009; 1791(7):628-35. DOI: 10.1016/j.bbalip.2009.02.008. View

Rindler M, Xu C, Gumper I, Smith N, Neubert T . Proteomic analysis of pancreatic zymogen granules: identification of new granule proteins. J Proteome Res. 2007; 6(8):2978-92. PMC: 2582026. DOI: 10.1021/pr0607029. View

Lopez-Marques R, Poulsen L, Hanisch S, Meffert K, Buch-Pedersen M, Jakobsen M . Intracellular targeting signals and lipid specificity determinants of the ALA/ALIS P4-ATPase complex reside in the catalytic ALA alpha-subunit. Mol Biol Cell. 2010; 21(5):791-801. PMC: 2828965. DOI: 10.1091/mbc.e09-08-0656. View

10.

Seppen J, Rijnberg M, Cooreman M, Oude Elferink R . Lentiviral vectors for efficient transduction of isolated primary quiescent hepatocytes. J Hepatol. 2002; 36(4):459-65. DOI: 10.1016/s0168-8278(01)00308-7. View

11.

Bryde S, Hennrich H, Verhulst P, Devaux P, Lenoir G, Holthuis J . CDC50 proteins are critical components of the human class-1 P4-ATPase transport machinery. J Biol Chem. 2010; 285(52):40562-72. PMC: 3003355. DOI: 10.1074/jbc.M110.139543. View

12.

Paulusma C, Oude Elferink R, Jansen P . Progressive familial intrahepatic cholestasis type 1. Semin Liver Dis. 2010; 30(2):117-24. DOI: 10.1055/s-0030-1253221. View

13.

Paulusma C, Rudi de Waart D, Kunne C, Mok K, Oude Elferink R . Activity of the bile salt export pump (ABCB11) is critically dependent on canalicular membrane cholesterol content. J Biol Chem. 2009; 284(15):9947-54. PMC: 2665118. DOI: 10.1074/jbc.M808667200. View

14.

Wang L, Beserra C, Garbers D . A novel aminophospholipid transporter exclusively expressed in spermatozoa is required for membrane lipid asymmetry and normal fertilization. Dev Biol. 2004; 267(1):203-15. DOI: 10.1016/j.ydbio.2003.11.004. View

15.

Muthusamy B, Natarajan P, Zhou X, Graham T . Linking phospholipid flippases to vesicle-mediated protein transport. Biochim Biophys Acta. 2009; 1791(7):612-9. PMC: 3770137. DOI: 10.1016/j.bbalip.2009.03.004. View

16.

Coleman J, Molday R . Critical role of the beta-subunit CDC50A in the stable expression, assembly, subcellular localization, and lipid transport activity of the P4-ATPase ATP8A2. J Biol Chem. 2011; 286(19):17205-16. PMC: 3089563. DOI: 10.1074/jbc.M111.229419. View

17.

Paulusma C, Groen A, Kunne C, Ho-Mok K, Spijkerboer A, Rudi de Waart D . Atp8b1 deficiency in mice reduces resistance of the canalicular membrane to hydrophobic bile salts and impairs bile salt transport. Hepatology. 2006; 44(1):195-204. DOI: 10.1002/hep.21212. View

18.

Paulusma C, Elferink R . The type 4 subfamily of P-type ATPases, putative aminophospholipid translocases with a role in human disease. Biochim Biophys Acta. 2005; 1741(1-2):11-24. DOI: 10.1016/j.bbadis.2005.04.006. View

19.

Siggs O, Schnabl B, Webb B, Beutler B . X-linked cholestasis in mouse due to mutations of the P4-ATPase ATP11C. Proc Natl Acad Sci U S A. 2011; 108(19):7890-5. PMC: 3093471. DOI: 10.1073/pnas.1104631108. View

20.

Holthuis J, Levine T . Lipid traffic: floppy drives and a superhighway. Nat Rev Mol Cell Biol. 2005; 6(3):209-20. DOI: 10.1038/nrm1591. View