Amino acid substitution in the lactose carrier protein with the use of amber suppressors.

Abstract:

:Five lacY mutants with amber stop codons at known positions were each placed into 12 different suppressor strains. The 60 amino acid substitutions obtained in this manner were tested for growth on lactose-minimal medium plates and for transport of lactose, melibiose, and thiomethylgalactoside. Most of the amino acid substitutions in the regions of the putative loops (between transmembrane alpha helices) resulted in a reasonable growth rate on lactose with moderate-to-good transport activity. In one strain (glycine substituted for Trp-10), abnormal sugar recognition was found. The substitution of proline for Trp-33 (in the region of the first alpha helix) showed no activity, while four additional substitutions (lysine, leucine, cysteine, and glutamic acid) showed low activity. Altered sugar specificity was observed when Trp-33 was replaced by serine, glutamine, tyrosine, alanine, histidine, or phenylalanine. It is concluded that Trp-33 may be involved directly or indirectly in sugar recognition.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Huang AM,Lee JI,King SC,Wilson TH

doi

10.1128/jb.174.16.5436-5441.1992

keywords:

subject

Has Abstract

pub_date

1992-08-01 00:00:00

pages

5436-41

issue

16

eissn

0021-9193

issn

1098-5530

journal_volume

174

pub_type

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