Journal: iScience

Article Title: Variant-specific interaction of kinectin 1 with the multi-tRNA synthetase complex regulates ER sheet organization

doi: 10.1016/j.isci.2025.114303

Figure Lengend Snippet: The V2-encoded region of KTN1 binds to the N-terminal region of QARS in the MSC (A) Amino acid sequence comparison for mouse and human KTN1. Sequence identity was calculated for each exon. Exons shaded in gray are relatively less conserved, with human exon 27 being absent in mice. (B) Immunoblot analysis of the indicated proteins in FLAG immunoprecipitates prepared from HEK293T cells expressing human KTN1-FLAG constructs containing the indicated combinations of V2, V3, and V4. Representative data from three independent experiments. (C) Location of alternative exons and the EEF1D binding domain in human KTN1 (1357 amino acids, UniProt ID Q86UP2-1). Truncated KTN1 variants used for in vitro binding assay are shown at the same scale. (D) Schematic workflow of in vitro binding assay using GST-tagged truncated KTN1 variants as bait. (E) Immunoblot analysis of in vitro binding assay using HeLa KTN1 KO cell lysates as prey. Representative data from two independent experiments. (F) Schematic depiction of full-length human QARS and truncated variants: N-terminal (QARS-N) and C-terminal (QARS-C). (G and H) Immunoblot analysis of in vitro binding assay using purified 6xHis-tagged QARS-N (G) or in vitro-translated 6xHis-tagged MSC components (H) as prey. Representative data from three independent experiments. (I) Summary of in vitro binding assays, demonstrating QARS-N as the binding site for KTN1 V2(+). See also .

Article Snippet: EEF1D , Santa Cruz Biotechnology , Cat# sc-393731; RRID: NA.

Techniques: Sequencing, Comparison, Western Blot, Expressing, Construct, Binding Assay, In Vitro, Purification

Journal: iScience

Article Title: Variant-specific interaction of kinectin 1 with the multi-tRNA synthetase complex regulates ER sheet organization

doi: 10.1016/j.isci.2025.114303

Figure Lengend Snippet: The V2-encoded region of KTN1 binds to the N-terminal region of QARS in the MSC (A) Amino acid sequence comparison for mouse and human KTN1. Sequence identity was calculated for each exon. Exons shaded in gray are relatively less conserved, with human exon 27 being absent in mice. (B) Immunoblot analysis of the indicated proteins in FLAG immunoprecipitates prepared from HEK293T cells expressing human KTN1-FLAG constructs containing the indicated combinations of V2, V3, and V4. Representative data from three independent experiments. (C) Location of alternative exons and the EEF1D binding domain in human KTN1 (1357 amino acids, UniProt ID Q86UP2-1). Truncated KTN1 variants used for in vitro binding assay are shown at the same scale. (D) Schematic workflow of in vitro binding assay using GST-tagged truncated KTN1 variants as bait. (E) Immunoblot analysis of in vitro binding assay using HeLa KTN1 KO cell lysates as prey. Representative data from two independent experiments. (F) Schematic depiction of full-length human QARS and truncated variants: N-terminal (QARS-N) and C-terminal (QARS-C). (G and H) Immunoblot analysis of in vitro binding assay using purified 6xHis-tagged QARS-N (G) or in vitro-translated 6xHis-tagged MSC components (H) as prey. Representative data from three independent experiments. (I) Summary of in vitro binding assays, demonstrating QARS-N as the binding site for KTN1 V2(+). See also .

Article Snippet: EEF1D , Proteintech , Cat# 10630-1-AP; RRID: AB_2230993.

Techniques: Sequencing, Comparison, Western Blot, Expressing, Construct, Binding Assay, In Vitro, Purification