Kinetics of motor protein translocation on single-stranded DNA.

نویسندگان

  • Christopher J Fischer
  • Lake Wooten
  • Eric J Tomko
  • Timothy M Lohman
چکیده

The translocation of nucleic acid motor proteins along DNA or RNA can be studied in ensemble experiments by monitoring either the kinetics of the arrival of the protein at a specific site on the nucleic acid filament (generally one end of the filament) or the kinetics of ATP hydrolysis by the motor protein during translocation. The pre-steady state kinetic data collected in ensemble experiments can be analyzed by simultaneous global non-linear least squares (NLLS) analysis using a simple sequential "n-step" mechanism to obtain estimates of the rate-limiting step(s) in the translocation cycle, the average "kinetic step-size," and the efficiency of coupling ATP binding and hydrolysis to translocation.

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

Detecting translocation of individual single stranded DNA homopolymers through a fabricated nanopore chip.

Fabricated solid-state nanopore chips are used to probe individual single stranded DNA (ssDNA) homopolymers. Based on the analysis of the current blockage caused by DNA translocation through a voltage-biased nanopore, we discovered that the hydrodynamic diameter of ssDNA homopolymer helix is comparable to that of double stranded DNA (dsDNA) helix. This proof-of-principle demonstration shows tha...

متن کامل

Challenges to Design and Develop of DNA Aptamers for Protein Targets. I. Optimization of Asymmetric PCR for Generation of a Single Stranded DNA Library

Aptamers, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as SELEX. In the amplification and regeneration step of SELEX technique, dsDNA is conversed to ssDNA. Asymmetric PCR is one of the methods used for the generation of ssDNA. The purpose of this study was to design a random DNA library for selection of aptamers wit...

متن کامل

Challenges to Design and Develop of DNA Aptamers for Protein Targets. I. Optimization of Asymmetric PCR for Generation of a Single Stranded DNA Library

Aptamers, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as SELEX. In the amplification and regeneration step of SELEX technique, dsDNA is conversed to ssDNA. Asymmetric PCR is one of the methods used for the generation of ssDNA. The purpose of this study was to design a random DNA library for selection of aptamers wit...

متن کامل

Stepwise translocation of nucleic acid motors.

Recent single molecule studies have made a significant contribution to the understanding of the molecular mechanism involved in the movement of motor proteins which process DNA and RNA. Measurement of stepsize in two disparate motors, NS3 helicase and ribosome both revealed 3-bp steps, which consist of three hidden substeps. Combined with previous structural studies, NS3 is likely taking a sing...

متن کامل

جداسازی پروتئین LMG از بافت کبد موش و میانکنش آن با

ABSTRACT In eukaryote cells, DNA is complexed with a series of basic proteins making units of chromatin structure named nucleosomes. In addition, nonhistone proteins with different function are the components of chromatin. Among these proteins, a group with a low mobility on gel electrophoresis have been identified and named LMG. In this study a LMG protein with a molecular weigh of 160 ...

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:
  • Methods in molecular biology

دوره 587  شماره 

صفحات  -

تاریخ انتشار 2010