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J Recept Signal Transduct Res

Title:		Investigation of growth hormone releasing hormone receptor structure and activity using yeast expression technologies
Author(s):		Kajkowski EM; Price LA; Pausch MH; Young KH; Ozenberger BA;
Address:		"American Cyanamid Company, Princeton, New Jersey 08543, USA"
Journal Title:		J Recept Signal Transduct Res
Year:		1997
Volume:		17
Issue:		1-Mar
Page Number:		293 - 303
DOI:		10.3109/10799899709036610
ISSN/ISBN:		1079-9893 (Print) 1079-9893 (Linking)
Abstract:		"Growth hormone releasing hormone (GHRH) is the positive regulator of growth hormone synthesis and secretion in the anterior pituitary. The peptide confers activity by binding to a seven transmembrane domain G protein-coupled receptor. Signal transduction proceeds through subsequent G alpha s stimulation of adenylyl cyclase. To investigate ligand/receptor and receptor/G protein associations, the human GHRH receptor was expressed in a modified S. cerevisiae strain which allows for facile measurement of receptor activity by cell prototrophy mediated by a reporter gene coupled to the yeast pheromone response pathway. GHRH-dependent signal activation in this system required the substitution of yeast G alpha protein with proteins containing C-terminal regions of G alpha s. A D60G variant (analogous to the little mouse mutation) of the receptor failed to respond to agonist. In parallel studies, GHRH29 and the N-terminal extracellular region of the receptor were expressed as Gal4 fusion proteins in a 2-hybrid assay. A specific interaction between these proteins was readily observed. The D60G mutation was engineered into the receptor fusion protein. This protein failed to interact with the ligand fusion, confirming the specificity of the association between unmodified proteins. These two yeast expression technologies should prove invaluable in additional structure/activity analyses of this ligand/receptor pair as well as other peptide ligands and receptors"
Keywords:		"Animals Fungal Proteins/genetics/metabolism *GTP-Binding Protein alpha Subunits GTP-Binding Protein alpha Subunits, Gq-G11 GTP-Binding Protein alpha Subunits, Gs/metabolism GTP-Binding Proteins/metabolism *Heterotrimeric GTP-Binding Proteins Humans Mice P;"
Notes:		"MedlineKajkowski, E M Price, L A Pausch, M H Young, K H Ozenberger, B A eng England 1997/01/01 J Recept Signal Transduct Res. 1997 Jan-May; 17(1-3):293-303. doi: 10.3109/10799899709036610"