Document Detail


The effect of net charge on the solubility, activity, and stability of ribonuclease Sa.
MedLine Citation:
PMID:  11369859     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The net charge and isoelectric pH (pI) of a protein depend on the content of ionizable groups and their pK values. Ribonuclease Sa (RNase Sa) is an acidic protein with a pI = 3.5 that contains no Lys residues. By replacing Asp and Glu residues on the surface of RNase Sa with Lys residues, we have created a 3K variant (D1K, D17K, E41K) with a pI = 6.4 and a 5K variant (3K + D25K, E74K) with a pI = 10.2. We show that pI values estimated using pK values based on model compound data can be in error by >1 pH unit, and suggest how the estimation can be improved. For RNase Sa and the 3K and 5K variants, the solubility, activity, and stability have been measured as a function of pH. We find that the pH of minimum solubility varies with the pI of the protein, but that the pH of maximum activity and the pH of maximum stability do not.
Authors:
K L Shaw; G R Grimsley; G I Yakovlev; A A Makarov; C N Pace
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Protein science : a publication of the Protein Society     Volume:  10     ISSN:  0961-8368     ISO Abbreviation:  Protein Sci.     Publication Date:  2001 Jun 
Date Detail:
Created Date:  2001-05-22     Completed Date:  2001-10-18     Revised Date:  2013-06-11    
Medline Journal Info:
Nlm Unique ID:  9211750     Medline TA:  Protein Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1206-15     Citation Subset:  IM    
Affiliation:
Department of Medical Biochemistry and Genetics, Texas A&M University, College Station, Texas 77843, USA.
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MeSH Terms
Descriptor/Qualifier:
Aspartic Acid / chemistry
Circular Dichroism
Escherichia coli / metabolism
Glutamic Acid / chemistry
Hydrogen-Ion Concentration
Isoenzymes / chemistry*
Kinetics
Lysine / chemistry
Models, Molecular
Mutagenesis, Site-Directed
Protein Conformation
Protein Denaturation
Ribonucleases / chemistry*
Solubility
Temperature
Thermodynamics
Grant Support
ID/Acronym/Agency:
GM 37039/GM/NIGMS NIH HHS; TW01058/TW/FIC NIH HHS
Chemical
Reg. No./Substance:
0/Isoenzymes; 56-84-8/Aspartic Acid; 56-86-0/Glutamic Acid; 56-87-1/Lysine; EC 3.1.-/Ribonucleases; EC 3.1.-/ribonuclease Sa3
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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