Bacterial immunoglobulin a proteases monitored by continuous Spectrophotometry

FEBS Letters

Reinholdt, Jesper¹  Bleeg, Henry S.¹  Kilian, Mogens¹ 

[1] Department of Oral Biology, Royal Dental College, Vennelyst Boulevard, DK-8000 Aarhus C, Denmark

关键词: Immunoglobulin A;    IgA protease;    Polyethylene glycol;    Virulence;    Haemophilus;   

DOI  :  10.1016/0014-5793(85)80402-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

IgA proteases were estimated in a turbid aqueous two-phase system with 10% polyethylene glycol-Tris buffer, where IgA spontaneously concentrates in microscopic spherical particles (< 1 μm). After enzymatic cleavage of IgA into Fab_α and Fc_α fragments, these fragments are soluble and decreasing turbidity is observed. The reaction may be followed by conventional spectrophotometry. In this manner, IgA proteases may be estimated in 10 min. Examples of the utility of the method are given with results from inhibitor studies, estimation of K_m and purification of IgA protease from Haemophilus influenzae.

【 授权许可】

Unknown