Isolation of a matrix that binds medial Golgi enzymes

Slusarewicz, Paul; Nilsson, Tommy; Hui, Norman; Watson, Rose; Warren, Graham

doi:10.1083/jcb.124.4.405

Cited by 176 publications

(122 citation statements)

References 48 publications

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“…The extraction of both GnTI and GnTII from rat liver is known to be salt-dependent (28,40). In this report, we have demonstrated that these enzymes behaved in a similar manner in transfected cell lines, indicating that this is an inherent property of these proteins.…”

Section: Discussionsupporting

confidence: 55%

“…These results contrast with both Gal-T1 and HT that were solubilized in low salt and remained in the supernatant of a high speed spin. Slusarewicz et al (40) have previously shown that the medial Golgi enzymes, GnTI and ␣-mannosidase II from rat liver membranes interact with a detergent-insoluble intercisternal matrix in the absence of NaCl; the interaction of GnTI with the insoluble matrix was fully dissociated with 50 -100 mM NaCl (40). We concur that GnTI is extracted from Golgi membranes in a NaCl-dependent manner but also observe that GnTI is solubilized as high molecular weight complexes, which dissociate in a salt-dependent manner, a behavior shared by GnTII.…”

Section: Discussionmentioning

confidence: 99%

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Medial Golgi but Not Late Golgi Glycosyltransferases Exist as High Molecular Weight Complexes

Opat¹,

Houghton

Gleeson

2000

Journal of Biological Chemistry

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To investigate the organization of Golgi glycosyltransferases and their mechanism of localization, we have compared the properties of a number of medial and late acting Golgi enzymes. The medial Golgi enzymes, Nacetylglucosaminyltransferase I and II (GnTI and Gn-TII) required high salt for solubilization and migrated as high molecular weight complexes on sucrose density gradients. In contrast, the late acting Golgi enzymes, ␤1,4-galactosyltransferase and ␣1,2-fucosyltransferase, were readily solubilized in low salt and migrated as monomers/dimers by sucrose density gradient centrifugation. Analysis of membrane-bound GnTI chimeras indicates that the formation of high molecular weight complexes does not require the transmembrane domain and cytoplasmic tail sequences of GnTI. Furthermore, a soluble form of GnTI, containing the stem region and catalytic domain, accumulated in the Golgi prior to secretion, in contrast to ␤1,4-galactosyltransferase. Soluble GnTI, which also associated with high molecular weight complexes, was comparable with membranebound GnTI in its ability to glycosylate newly synthesized glycoproteins in vivo. Mutation of charged residues within the stem region of GnTI, known to be important for "kin recognition", had no effect on the efficiency of Golgi localization, the inclusion into high molecular weight complexes, nor functional activity in vivo. The differences in behavior between the medial and late acting Golgi enzymes may contribute to their differential localization and their ability to glycosylate efficiently in the correct Golgi subcompartment.

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Section: Discussionsupporting

confidence: 55%

Section: Discussionmentioning

confidence: 99%

Medial Golgi but Not Late Golgi Glycosyltransferases Exist as High Molecular Weight Complexes

Opat¹,

Houghton

Gleeson

2000

Journal of Biological Chemistry

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“…5B,D). GM130 protein was originally purified from Golgi membranes (Nakamura et al, 1995) and identified in the stacked cisternae of the Golgi complex (Slusarewicz et al, 1994). The MACF1b antibody staining near the nucleus colocalized with that of GM130 (compare A with B and C with D of Fig.…”

Section: Macf1b Localizes To the Golgi Complexmentioning

confidence: 99%

Microtubule actin crosslinking factor 1b: a novel plakin that localizes to the Golgi complex

Lin

Chen

Leung

et al. 2005

Journal of Cell Science

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MACF1 (microtubule actin crosslinking factor), also called ACF7 (actin crosslinking family 7) is a cytoskeletal linker protein that can associate with both actin filaments and microtubules. We have identified a novel alternatively spliced isoform of MACF1. We named this isoform MACF1b and renamed the original isoform MACF1a. MACF1b is identical to MACF1a, except that it has a region containing plakin (or plectin) repeats in the middle of the molecule. MACF1b is ubiquitously expressed in adult tissues with especially high levels in the lung. We studied the subcellular localization of MACF1b proteins in mammalian cell lines. In two lung cell lines, MACF1b was chiefly localized to the Golgi complex. Upon treatments that disrupt the Golgi complex, MACF1b redistributed into the cytosol, but remained co-localized with the dispersed Golgi ministacks. MACF1b proteins can be detected in the enriched Golgi fraction by western blotting. The domain of MACF1b that targets it to the Golgi was found at the N-terminal part of the region that contains the plakin repeats. Reducing the level of MACF1 proteins by small-interfering RNA resulted in the dispersal of the Golgi complex.

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“…It is known that there are proteinaceous links that hold apposed cisternae together (16). The Golgi matrix is a detergent and salt resistant complex that is easily isolated from purified rat liver Golgi membranes (17). The Golgi matrix specifically binds the cytosolic region of Golgi resident transmembrane proteins, so it is supposed to contain intercisternal structural components that support the flattened and stacked cisternal structure.…”

Section: Gm130 As a Component Of The Golgi Matrixmentioning

confidence: 99%

Emerging New Roles of GM130, a cis-Golgi Matrix Protein, in Higher Order Cell Functions

2010

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Abstract. GM130 is a peripheral membrane protein strongly attached to the Golgi membrane and is isolated from the detergent and salt resistant Golgi matrix. GM130 is rich in coiled-coil structures and predicted to take a rod-like shape. Together with p115, giantin, and GRASP65, GM130 facilitates vesicle fusion to the Golgi membrane as a vesicle "tethering factor". GM130 is also involved in the maintenance of the Golgi structure and plays a major role in the disassembly and reassembly of the Golgi apparatus during mitosis. Emerging evidence suggests that GM130 is involved in the control of glycosylation, cell cycle progression, and higher order cell functions such as cell polarization and directed cell migration. This creates the potential for novel Golgitargeted drugs and treatments for various diseases including glycosylation defects, immune diseases, and cancer.

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Isolation of a matrix that binds medial Golgi enzymes

Abstract: Abstract. Rat liver Golgi stacks were extracted with

Cited by 176 publications

References 48 publications

Medial Golgi but Not Late Golgi Glycosyltransferases Exist as High Molecular Weight Complexes

Medial Golgi but Not Late Golgi Glycosyltransferases Exist as High Molecular Weight Complexes

Microtubule actin crosslinking factor 1b: a novel plakin that localizes to the Golgi complex

Emerging New Roles of GM130, a cis-Golgi Matrix Protein, in Higher Order Cell Functions

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