Plaué S, Muller S, van Regenmortel M H
Institut de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Strasbourg, France.
J Exp Med. 1989 May 1;169(5):1607-17. doi: 10.1084/jem.169.5.1607.
Two peptides of eight (T2) and 10 (T1) residues corresponding to the branched moiety of ubiquitinated histone H2A have been synthesized and used for raising specific antibodies in rabbits. Antisera to peptide T1 reacted in ELISA with T1 and with H2A but not with ubiquitin; antisera to peptide T2 reacted with T2 but not with H2A or ubiquitin. When tested in immunoblotting, both peptide antisera reacted with ubiquitinated H2A but not with unconjugated H2A or with ubiquitin. Sera from patients with systemic lupus erythematosus (SLE) were shown previously to react with ubiquitin in ELISA and immunoblotting. When tested for their ability to react in ELISA with synthetic peptides T1 and T2, 96% of the SLE sera (diluted 1:500) that recognized ubiquitin also reacted with peptide T2. Of the SLE sera that did not react with ubiquitin, only 13% possessed antibodies able to bind peptide T2. Antibodies from seven SLE sera, purified on a T2-immunoadsorbent column, were also able to react either with H2A, and in three cases also with ubiquitin.
合成了与泛素化组蛋白H2A的分支部分相对应的8个残基(T2)和10个残基(T1)的两种肽,并用于在兔体内产生特异性抗体。肽T1的抗血清在ELISA中与T1和H2A反应,但不与泛素反应;肽T2的抗血清与T2反应,但不与H2A或泛素反应。在免疫印迹试验中,两种肽抗血清均与泛素化H2A反应,但不与未结合的H2A或泛素反应。先前已证明系统性红斑狼疮(SLE)患者的血清在ELISA和免疫印迹中与泛素反应。当检测它们在ELISA中与合成肽T1和T2反应 的能力时,96%识别泛素的SLE血清(稀释1:500)也与肽T2反应。在不与泛素反应的SLE血清中,只有13%具有能够结合肽T2的抗体。在T2免疫吸附柱上纯化的来自7例SLE血清的抗体也能够与H2A反应,在3例中还能与泛素反应。