A universal flow cytometry assay for screening carbohydrate-active enzymes using glycan microspheres.

نویسندگان

  • Aarthi Chandrasekaran
  • Kai Deng
  • Chung-Yan Koh
  • Taichi Takasuka
  • Lai F Bergeman
  • Brian G Fox
  • Paul D Adams
  • Anup K Singh
چکیده

We describe a simple, multiplexed assay that integrates glycan synthesis, bioconjugation to microspheres, fluorescent chemical/biochemical detection and multiparameter flow cytometric analysis to screen activities of different families of carbohydrate-active enzymes.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Correlation Index-Based Responsible-Enzyme Gene Screening (CIRES), a Novel DNA Microarray-Based Method for Enzyme Gene Involved in Glycan Biosynthesis

BACKGROUND Glycan biosynthesis occurs though a multi-step process that requires a variety of enzymes ranging from glycosyltransferases to those involved in cytosolic sugar metabolism. In many cases, glycan biosynthesis follows a glycan-specific, linear pathway. As glycosyltransferases are generally regulated at the level of transcription, assessing the overall transcriptional profile for glycan...

متن کامل

Quinoxalinone Inhibitors of the Lectin DC-SIGN.

The C-type lectin dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) can serve as a docking site for pathogens on the surface of dendritic cells. Pathogen binding to DC-SIGN can have diverse consequences for the host. DC-SIGN can facilitate HIV-1 dissemination, but the interaction of Mycobacterium tuberculosis with DC-SIGN is important for host immunity. Th...

متن کامل

Glycan Sequencing Using Exoglycosidases

An increasing number of structurally related families of enzymes that degrade, modify, or create glycosidic bonds are being identified and characterized. Davies, et al., estimated that carbohydrate-active enzymes represent 1‐3% of all proteins encoded by the genomes of most organisms, and identification of 12,000+ open reading frames for glycoside hydrolases and glycosyltransferases was project...

متن کامل

Multiplex suspension array for human anti-carbohydrate antibody profiling.

Glycan-binding antibodies form a significant subpopulation of both natural and acquired antibodies and play an important role in various immune processes. They are for example involved in innate immune responses, cancer, autoimmune diseases, and neurological disorders. In the present study, a microsphere-based flow-cytometric immunoassay (suspension array) was applied for multiplexed detection ...

متن کامل

Remodeling of glycans using glycosyltransferase genes.

Remodeling of glycans on the cell surface is an essential technique to analyze cellular function of lectin-glycan ligand interaction. Here we describe the methods to identify the responsible enzyme (glycosyltransferase) regulating the expression of the glycan of interest and to modulate the glycan expression by overexpressing the glycosyltransferase gene. For the identification of the responsib...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Chemical communications

دوره 49 48  شماره 

صفحات  -

تاریخ انتشار 2013