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) |