The Mammalian Calcium-binding Protein, Nucleobindin (CALNUC), Is a Golgi Resident Protein

doi:10.1083/jcb.141.7.1515

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J. Cell Biol., Volume 141, Number 7, June 29, 1998 1515-1527

The Mammalian Calcium-binding Protein, Nucleobindin (CALNUC), Is a Golgi Resident Protein

Ping Lin, Helen Le-Niculescu, Robert Hofmeister, J. Michael McCaffery, Mingjie Jin, Hanjo Hennemann, Tammie McQuistan, Luc De Vries, and Marilyn Gist Farquhar

Division of Cellular and Molecular Medicine and Department of Pathology, University of California, San Diego, La Jolla, California 92093-0651

We have identified CALNUC, an EF-hand, Ca²⁺-binding protein, as a Golgi resident protein. CALNUC corresponds to a previouslyidentified EF-hand/calcium-binding protein known as nucleobindin.CALNUC interacts with G $alpha$ _i3 subunits in the yeast two-hybrid systemand in GST-CALNUC pull-down assays. Analysis of deletion mutantsdemonstrated that the EF-hand and intervening acidic regions arethe site of CALNUC's interaction with G $alpha$ _i3. CALNUC is found inboth cytosolic and membrane fractions. The membrane pool is tightlyassociated with the luminal surface of Golgi membranes. CALNUCis widely expressed, as it is detected by immunofluorescence inthe Golgi region of all tissues and cell lines examined. By immunoelectronmicroscopy, CALNUC is localized to cis-Golgi cisternae and thecis-Golgi network (CGN). CALNUC is the major Ca²⁺-binding protein detected by ⁴⁵Ca²⁺-binding assay on Golgi fractions. The properties of CALNUC andits high homology to calreticulin suggest that it may play a keyrole in calcium homeostasis in the CGN and cis-Golgi cisternae.

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