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PMID: 9600905 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Two amino acid substitutions convert a guanylyl cyclase, RetGC-1, into an adenylyl cyclase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 95 ·No. 11 ·1998-05-26 ·Pages 5993-7

Tucker CL, Hurley JH, Miller TR, Hurley JB

Abstract

Guanylyl cyclases (GCs) and adenylyl cyclases (ACs) have fundamental roles in a wide range of cellular processes. Whereas GCs use GTP as a substrate to form cGMP, ACs catalyze the analogous conversion of ATP to cAMP. Previously, a model based on the structure of adenylate cyclase was used to predict the structure of the nucleotide-binding pocket of a membrane guanylyl cyclase, RetGC-1. Based on this model, we replaced specific amino acids in the guanine-binding pocket of GC with their counterparts from AC. A change of two amino acids, E925K together with C995D, is sufficient to completely alter the nucleotide specificity from GTP to ATP. These experiments strongly validate the AC-derived RetGC-1 structural model and functionally confirm the role of these residues in nucleotide discrimination.

MeSH Terms
Adenylyl Cyclases/chemistry,genetics Amino Acid Sequence Amino Acid Substitution Binding Sites/genetics Cell Line DNA Mutational Analysis Guanylate Cyclase/chemistry,genetics Humans Membrane Proteins/chemistry,genetics Models, Molecular Molecular Sequence Data Structure-Activity Relationship Substrate Specificity/genetics
Chemicals
Membrane Proteins Adenylyl Cyclases Guanylate Cyclase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tucker C L
Howard Hughes Medical Institute and Department of Biochemistry, Box 357370, University of Washington, Seattle, WA 98195, USA.
Hurley J H
Miller T R
Hurley J B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.95.11.5993
Published
1998-05-26
Pages
5993-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC27573
Subset
IM
Grants
NEI NIH HHS · F32 EY006641 · United States
NEI NIH HHS · R01 EY006641 · United States
NEI NIH HHS · EY06641 · United States
External Links
PubMed source DOI: 10.1073/pnas.95.11.5993 PMC Full Text View journal details
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