RRC ID 1497
Author Nakanishi K, Komatsu Y, Kogawa N, Matsushita H.
Title Analysis of eluted peptides from type 1 diabetes-susceptible HLA class II molecules identified novel islet protein, heparin/heparan sulfate-interacting protein.
Journal Biochem Biophys Res Commun
Abstract Identification of peptides derived from pancreatic islet and presented by type 1 diabetes-susceptible MHC class II molecules has great significance to elucidate the pathogenesis of type 1 diabetes. A bulk culture of Epstein-Barr virus-transformed B-cells, which were established from a 22-year-old type 1 diabetic woman with HLA-DR4 and -DQw8, was pulsed with the homogenate of a human embryonic pancreas-derived cell line 1B2C6, and another culture was not pulsed with antigen. Peptide fractions were obtained by treatment of affinity-purified HLA-DR and -DQ molecules with 0.1% trifluoroacetic acid, and were subjected to reverse-phase high performance liquid chromatography (RP-HPLC). The RP-HPLC profiles of peptides derived from DR molecules revealed three peaks that specifically appeared after pulsing, but no such peaks were obtained from DQ molecules. From one of these three peaks, a peptide that consisted of 14 amino acids (AKSXNHTXXNQXRK, where X represents the undetermined amino acids) was identified. This peptide was derived from heparin/heparan sulfate-interacting protein (HIP). Immunostaining of pancreatic sections using antiserum for HIP peptide revealed exclusive staining of the islets. Thus, HIP was identified as an islet protein naturally processed and presented by HLA-DR4 molecules.
Volume 329(1)
Pages 356-61
Published 2005-4-1
DOI 10.1016/j.bbrc.2005.01.144
PII S0006-291X(05)00195-6
PMID 15721314
MeSH Alleles Amino Acids / chemistry B-Lymphocytes / cytology Blood Coagulation Factors / chemistry* Blood Coagulation Factors / physiology* Blotting, Western Cell Line Cells, Cultured Chromatography, High Pressure Liquid Diabetes Mellitus, Type 1 / immunology* Female Genes, MHC Class II* HLA-DQ Antigens / chemistry HLA-DR Antigens / chemistry HLA-DR4 Antigen / chemistry Heparin / metabolism* Heparitin Sulfate / chemistry* Herpesvirus 4, Human / metabolism Humans Immunohistochemistry Islets of Langerhans / cytology* Islets of Langerhans / metabolism* Leukocytes, Mononuclear / metabolism Pancreas / embryology* Pancreas / metabolism Peptides / chemistry* Protein Binding RNA-Binding Proteins Ribosomal Proteins Time Factors Trifluoroacetic Acid / metabolism
IF 2.985
Times Cited 4
WOS Category BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells 1B2C6(RCB0795)