SCIENCE@home

Saturation Mutagenesis of Human Interleukin-3 (*)

P. O. Olins,S. C. Bauer,S. Braford-Goldberg,K. Sterbenz,J. Polazzi,M. Caparon,Barbara K. Klein,A. Easton,Kumnan Paik,Jon A. Klover,B. Thiele,J. Mckearn

Published 1995 in Journal of Biological Chemistry

ABSTRACT

A deletion variant of human interleukin-3, hIL-3, was produced in the periplasmic space of Escherichia coli and had full activity in an AML193.1.3 cell proliferation assay. Libraries of random single-amino acid substitutions were constructed at each of 105 positions in the gene for hIL-3. Approximately eight single-site substitutions at each position were produced in osmotic shock fractions and screened for activity. 15 mutants were found with bioactivity of 5-26-fold greater than that of native hIL-3. The majority of amino acids in hIL-3 could be substituted without substantial loss of activity. Substitution of residues predicted to be in the hydrophobic core of the protein often resulted in reduced activity and/or low accumulation levels. Only five residues predicted to be on the surface of the protein were intolerant of substitution and hence are candidates for sites of interaction with the receptor. We therefore propose that the majority of residues in hIL-3 serve a structural role and permit the display of a few key residues in the correct configuration for recognition by the receptor.

PUBLICATION RECORD

CITATION MAP

EXTRACTION MAP

CLAIMS

  • No claims are published for this paper.

CONCEPTS

  • No concepts are published for this paper.

REFERENCES

Showing 1-60 of 60 references · Page 1 of 1

CITED BY

Showing 1-45 of 45 citing papers · Page 1 of 1