Development of process for in-vitro protein synthesis from living cells followed by dissociation of ribosomes into subunits is discussed. Process depends on dialysis or use of chelating agents. Operation of process and advantages over previous methods are outlined

Friedman, M.

Lu, P.

Rich, A.

English

NASA Technical Reports Server

October 1972	 B72-10653 
S NASA TECH BRIEF 
NASA Headquarters
	 -4r 
NASA Tech Briefs announce new technology derived from the U.S. space program. They are issued to encourage 
commercial application. Tech Briefs are available on a subscription basis from the National Technical Information 
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be directed to the Technology Utilization Office, NASA, Code KT, Washington, D.C. 20546. 
A Process Yields Large Quantities of Pure Ribosome Subunits 
S
The problem: 
Ribosome subunits which are used in the study of their 
properties or for in-vitro protein synthesis are obtained by 
isolation of ribosomes from the living cells followed by 
dissociation of these ribosomes into subunits by either 
dialysis or with chelating agents. This process is inefficient 
because it yields a large quantity of impure subunits, or 
the so called "derived" subunits, which contain partially 
completed proteins that were being synthesized when the 
cell was broken. The yield of "native" subunits or the 
subunits which are about to initiate a new round of 
protein synthesis is unfortunately very small. 
The solution: 
A new process was developed which improves the yield 
of "native" subunits to 90. This process was applied to 
E. coli. 
How it's done: 
Before the cells are broken, they are cooled to below 
the critical temperature so that ribosomes continue 
protein synthesis but cannot reinitiate a new cycle. The 
result is a large accumulation of "native" subunits which 
are then isolated in the conventional way without 
disassembly of the ribosome itself.
The advantages of the presented method are: 1) a 
lengthy or expensive step in the preparation is omitted; 
2) the subunits are free of contaminating partially-
completed protein; 3) the product is in its naturally active 
form; 4) the product is obtained in high yield; and 5) the 
method is not restricted to particular mutants but can be 
applied to all cells. 
Note: 
Requests for further information may be directed to: 
Technology Utilization Officer 
NASA Headquarters 
Code KT 
Washington, D.C. 20546 
Reference: B72-10653 
Patent status: 
NASA has decided not to apply for a patent. 
Source: M. Friedman, P. Lu, and

A. Rich of
Massachusetts Institute of Technology

under contract to

NASA Headquarters
(HQN-1 0662) 
I Category 05 
This document was prepared under the sponsorship of the National
	 Government assumes any liability resulting from the use of the 
Aeronautics and Space Administration. Neither the United States
	 information contained in this document, or warrants that such use 
Government nor any person acting on behalf of the United States
	 will be free from privately owned rights.
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A process yields large quantities of pure ribosome subunits

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