Development and application of antibody against phosphohistidine.

2013 Fiscal Year Final Research Report

• Project/Area Number: 24659138
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: General medical chemistry
• Research Institution: Kurume University
• Principal Investigator: MIURA YOSHIKI 久留米大学, 分子生命科学研究所, 講師 (90279240)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: リン酸化ヒスチジン / 抗体

Research Abstract

Histidine residue is one of the major site of protein phosphorylation. Raising antibodies against phosphohistidine has been difficult due to its chemically labile nature. Using chemically stable phosphohistidine analogue, we developed mouse monoclonal antibodies against phosphohistidine. However, specificity of these antibodies were alone insufficient to identify phosphohistidine, they also react but weakly with phosphoserine, phosphothreonine and phosphotyrosine. On the other hand, polyclonal antibodies raised in rabbit were highly specific to phosphohistidine. After affinity purification using phosphohistidine column, purified antibodies only recognized phosphohistidine.