Structural and Biochemical Characterization of Cysteinylation in Broadly Neutralizing Antibodies to HIV-1.
Structural and Biochemical Characterization of Cysteinylation in Broadly Neutralizing Antibodies to HIV-1.
复制标题
DOI：
10.1016/j.jmb.2021.167303
复制
发表时间：
2021-12-03
影响因子：
5.6
通讯作者：
Wilson IA
中科院分区：
文献类型：
--
作者：
Omorodion O;Wilson IA
半胱氨酸化是抗体中很少研究的翻译后修饰。在针对HIV-1的bnAb谱系中鉴定了半胱氨酸化。半胱氨酸化修饰在晶体结构中清晰可见。与以前的研究相比，半胱氨酸化不损害与抗原的结合。这些发现为抗体半胱氨酸化的作用提供了进一步的观点。在自然HIV-1感染过程中，某些个体体内产生了具有特殊广度和效力的抗体。因此，广泛中和抗体（bnAb）的诱导和亲和力成熟是HIV-1疫苗开发的中心目标。bnAb的功能特性也使它们作为免疫抑制剂具有吸引力，这导致它们的生产和优化用于被动免疫治疗。该过程需要体外制造和监测任何异质表达，特别是当产生具有不同生物活性水平的抗体亚群时。抗体的翻译后修饰（PTM）可导致异质性，是本研究的重点。具体而言，我们已经研究了半胱氨酸化的bnAb谱系（PCDN家族）靶向N332-聚糖supersite的表面包膜糖蛋白（Env）的HIV-1。该PTM通过用分子半胱氨酸封端未配对的半胱氨酸残基来定义。通过色谱法和质谱法，我们能够表征在哺乳动物细胞中表达时半胱氨酸化和非半胱氨酸化抗体的亚群。两种PCDN抗体的晶体结构代表半胱氨酸化抗体的第一结构，并揭示在这种情况下半胱氨酸化位于CDRH 3中。生物物理学研究表明，这些HIV-1抗体的半胱氨酸化不干扰抗原结合，据报道，这发生在其他半胱氨酸化抗体中。因此，这些研究强调需要进一步研究抗HIV和其他bnAb中的半胱氨酸化。
Cysteinylation is a little-studied post-translational modification in antibodies. Cysteinylation was identified in a lineage of bnAbs to HIV-1. The cysteinylation modification is clearly visualized in the crystal structures. Cysteinylation does not impair binding to antigen, compared to previous studies. These findings provide further perspectives on the role of antibody cysteinylation. Antibodies with exceptional breadth and potency have been elicited in some individuals during natural HIV-1 infection. Elicitation and affinity maturation of broadly neutralizing antibodies (bnAbs) is therefore the central goal of HIV-1 vaccine development. The functional properties of bnAbs also make them attractive as immunotherapeutic agents, which has led to their production and optimization for passive immunotherapy. This process requires in vitro manufacturing and monitoring of any heterogeneous expression, especially when subpopulations of antibodies are produced with varying levels of biological activity. Post-translational modification (PTM) of antibodies can contribute to heterogeneity and is the focus of this study. Specifically, we have investigated cysteinylation in a bnAb lineage (PCDN family) targeting the N332-glycan supersite on the surface envelope glycoprotein (Env) of HIV-1. This PTM is defined by capping of unpaired cysteine residues with molecular cysteine. Through chromatography and mass spectrometry, we were able to characterize subpopulations of cysteinylated and non-cysteinylated antibodies when expressed in mammalian cells. The crystal structures of two PCDN antibodies represent the first structures of a cysteinylated antibody and reveal that the cysteinylation in this case is located in CDRH3. Biophysical studies indicate that cysteinylation of these HIV-1 antibodies does not interfere with antigen binding, which has been reported to occur in other cysteinylated antibodies. As such, these studies highlight the need for further investigation of cysteinylation in anti-HIV and other bnAbs.
登录
查看更多内容
影响因子：
2.9
作者：
Mahomed S;Garrett N;Karim QA;Zuma NY;Capparelli E;Baxter C;Gengiah T;Archary D;Samsunder N;Doria-Rose N;Moore P;Williamson C;Barouch DH;Fast PE;Pozzetto B;Hankins C;Carlton K;Ledgerwood J;Morris L;Mascola J;Abdool Karim S
通讯作者：
Abdool Karim S
影响因子：
6.7
作者：
Sanders RW;Derking R;Cupo A;Julien JP;Yasmeen A;de Val N;Kim HJ;Blattner C;de la Peña AT;Korzun J;Golabek M;de Los Reyes K;Ketas TJ;van Gils MJ;King CR;Wilson IA;Ward AB;Klasse PJ;Moore JP
通讯作者：
Moore JP
FC修饰的HIV-1人单克隆抗体VRC01LS的安全性和药代动力学：健康成年人中的1期开放标签临床试验。
影响因子：
15.8
作者：
Gaudinski MR;Coates EE;Houser KV;Chen GL;Yamshchikov G;Saunders JG;Holman LA;Gordon I;Plummer S;Hendel CS;Conan-Cibotti M;Lorenzo MG;Sitar S;Carlton K;Laurencot C;Bailer RT;Narpala S;McDermott AB;Namboodiri AM;Pandey JP;Schwartz RM;Hu Z;Koup RA;Capparelli E;Graham BS;Mascola JR;Ledgerwood JE;VRC 606 Study Team
通讯作者：
VRC 606 Study Team
影响因子：
14.9
作者：
Burley SK;Bhikadiya C;Bi C;Bittrich S;Chen L;Crichlow GV;Christie CH;Dalenberg K;Di Costanzo L;Duarte JM;Dutta S;Feng Z;Ganesan S;Goodsell DS;Ghosh S;Green RK;Guranović V;Guzenko D;Hudson BP;Lawson CL;Liang Y;Lowe R;Namkoong H;Peisach E;Persikova I;Randle C;Rose A;Rose Y;Sali A;Segura J;Sekharan M;Shao C;Tao YP;Voigt M;Westbrook JD;Young JY;Zardecki C;Zhuravleva M
通讯作者：
Zhuravleva M
影响因子：
64.5
作者：
Garces F;Sok D;Kong L;McBride R;Kim HJ;Saye-Francisco KF;Julien JP;Hua Y;Cupo A;Moore JP;Paulson JC;Ward AB;Burton DR;Wilson IA
通讯作者：
Wilson IA