Defining a novel 75-kDa phosphoprotein associated with SS-A/Ro and identification of distinct human autoantibodies

Wang, Dunrui; Buyon, Jill P.; Zhu, Weiguo; Chan, Edward K. L.

doi:10.1172/jci8003

Cited by 37 publications

(26 citation statements)

References 33 publications

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“…AB020664) identified by Nagase et al (27) contained an open reading frame that encoded a 733-amino acid protein; however, when the first methionine (with appropriate Kozak sequence (Ref. 37)) was used as the start site, the openreading frame encoded a protein of 653 amino acids (38). This protein's C terminus (amino acids 490 -653) was identical to and in frame with the Gaf-1 clone recovered in our screen.…”

Section: Identification Of ␥-Snap-bindingmentioning

confidence: 60%

“…Consistently, immunoprecipitation experiments show that ␥-SNAP is the major Gaf-1-associated protein in HepG2 cells. Gaf-1 is ubiquitously expressed in mammalian tissues based on Northern (27,38) and Western blotting analysis (Fig. 3).…”

Section: Discussionmentioning

confidence: 99%

“…Although the N-terminal domain may be significant for Gaf-1 binding to membranes (see below), no ATP binding has been detected (data not shown). Gaf-1 appears to be identical to a protein called pp75, which was identified in a two-hybrid screen using the SS-A/Ro autoantigen as bait (38).…”

Section: Identification Of ␥-Snap-bindingmentioning

confidence: 99%

“…Gaf-1 Distribution and Localization-Northern blot analysis by Nagase et al (27) and Wang et al (38) suggested that Gaf-1 was ubiquitously expressed in mammalian tissues as is ␥-SNAP (13). To confirm this we used an antibody against the C-terminal domain of Gaf-1 to probe extracts from various mouse tissues and cell lines.…”

Section: Identification Of ␥-Snap-bindingmentioning

confidence: 99%

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Gaf-1, a γ-SNAP-binding Protein Associated with the Mitochondria

Chen

et al. 2001

Journal of Biological Chemistry

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The role of ␣/␤-SNAP (Soluble NSF Attachment Protein) in vesicular trafficking is well established; however, the function of the ubiquitously expressed ␥-SNAP remains unclear. To further characterize the cellular role of this enigmatic protein, a two-hybrid screen was used to identify new, ␥-SNAP-binding proteins and to uncover potentially novel functions for ␥-SNAP. One such SNAP-binding protein, termed Gaf-1 (␥-SNAP associate factor-1) specifically binds ␥-but not ␣-SNAP. The full-length Gaf-1 (75 kDa) is ubiquitously expressed and is found stoichiometrically associated with ␥-SNAP in cellular extracts. This binding is distinct from other SNAP interactions since no ␣-SNAP or NSF coprecipitated with Gaf-1. Subcellular fractionation and immunofluorescence analysis show that Gaf-1 is peripherally associated with the outer mitochondrial membrane. Only a fraction of ␥-SNAP was mitochondrial with the balance being either cytosolic or associated with other membrane fractions. GFP-␥-SNAP and the C-terminal domain of Gaf-1 both show a reticular distribution in HEK-293 cells. This reticular structure colocalizes with Gaf-1 and mitochondria as well as with microtubules but not with other cytoskeletal elements. These data identify a class of ␥-SNAP interactions that is distinct from other members of the SNAP family and point to a potential role for ␥-SNAP in mitochondrial dynamics.The soluble N-ethylmaleimide-sensitive factor (NSF) 1 attachment protein family (␣-, ␤-, and ␥-SNAP) is thought to function in vesicular transport by mediating the binding of the NSF to the SNAP receptors (SNAREs) (reviewed in Refs.

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Section: Identification Of ␥-Snap-bindingmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 99%

Section: Identification Of ␥-Snap-bindingmentioning

confidence: 99%

Section: Identification Of ␥-Snap-bindingmentioning

confidence: 99%

See 2 more Smart Citations

Gaf-1, a γ-SNAP-binding Protein Associated with the Mitochondria

Chen

et al. 2001

Journal of Biological Chemistry

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“…In vitro translated proteins were examined for reactivity by sera using IP described (Chan et al, 1991;Wang et al, 1999). Controls included an unrelated phosphoprotein pp75 (Wang et al, 1999) produced at TnT products and mixed with a p90 TnT fragment to monitor the specificity of the IP reaction.…”

Section: Immunoprecipitation (Ip) Of Tnt Productsmentioning

confidence: 99%