In-cell covalent labeling of reactive His-tag fused proteins.
نویسندگان
چکیده
A new method for in-cell protein labeling was developed. This method employed a binding-induced nucleophilic reaction between the Cys-appended His-tag and the Ni(II)-NTA containing an α-chloroacetamide. Using this method, not only labeling of His-tag fused proteins but also the detection of a protein-protein interaction was achieved inside living cells.
منابع مشابه
Hydrazide reactive peptide tags for site-specific protein labeling.
New site-specific protein labeling (SSPL) reactions for targeting-specific, short peptides could be useful for the real-time detection of proteins inside of living cells. One SSPL approach matches bioorthogonal reagents with complementary peptides. Here, hydrazide reactive peptides were selected from phage-displayed libraries using reaction-based selections. Selection conditions included washes...
متن کامل[Development of specific protein labeling method by using small molecular probes].
Selective protein labeling with a small molecular probe is a versatile method for elucidating protein functions in living cells. In this review, we report a new protein labeling method that enables selective covalent modification of a tag-fused protein with small functional molecules. This method utilizes the specific interaction and rapid reaction between a short peptide tag and a molecular pr...
متن کاملHaloTag7: a genetically engineered tag that enhances bacterial expression of soluble proteins and improves protein purification.
Over-expression and purification of soluble and functional proteins remain critical challenges for many aspects of biomolecular research. To address this, we have developed a novel protein tag, HaloTag7, engineered to enhance expression and solubility of recombinant proteins and to provide efficient protein purification coupled with tag removal. HaloTag7 was designed to bind rapidly and covalen...
متن کاملهمسانهسازی و بیان ایمونوتوکسین اونتاک به صورت هیبریدی با دنباله اینتئینی
Introduction: Inteins (INT) are internal parts of a number of proteins in yeast and some other unicellular eukaryotes, which can be separated from the immature protein during protein splicing process. After identifying the mechanism of intein action, applications of these sequences are be considered in the single- step purification of recombinant proteins and different intein tags were develope...
متن کاملFluorescent substrates for covalent protein labeling catalyzed by microbial transglutaminase.
Novel small substrates with a variety of fluorophores were designed for the covalent labeling of proteins catalyzed by microbial transglutaminase (MTG). The new design is based on the flexibility in the substrate recognition of MTG for the substitution of the N-terminal protecting group of a conventional transglutaminase substrate, benzyloxycarbonyl-L-glutaminylglycine (Z-QG). Here we report fo...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Chemical communications
دوره 49 44 شماره
صفحات -
تاریخ انتشار 2013