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Much evidence indicates that ANG II may also participate in the development of pathological states. 3,4 The potential role of ANG II as a growth factor led to its implication in the pathogenesis of hypertension and atherosclerosis. 5 Bernard Lassegue, Kathy K. Griendling, and R. Wayne Alexander· Cardiology Division, Emory University School of Medicine, Atlanta, Georgia 30322. Angiotensin Receptors, edited by Juan M. M. Timmermans. Plenum Press, New York, 1994. 17 18 Bernard Lassegue et al. In order to elucidate the pathways underlying the striking multiplicity of effects of this single octapeptide hormone, many biochemical studies endeavored to isolate ANG II receptors, the cellular effectors that bind ANG II and transduce its physiological effects, and to dissect their signaling mechanisms.

TISSUE DISTRIBUTION OF THE AT1A AND AT1s SUBTYPES IN THE RAT To determine possible functional differences between these receptors, relative tissue distributions were examined. Since there is no notable pharmacological difference between the subtypes, mRNA expression has been measured. However, since ATJ probes used in previous studies hybridized to the nearly identical coding regions of ATIA and ATlB, the strategy was adopted of using PCR amplification of the untranslated divergent portions of the two subtypes.

The first report concentrated on mutations that may affect ANG II binding. 55 The replacement of any of the four cysteine residues from the extracellular domains by glycine markedly decreased affinity for [125I]-ANG II. This confirms the results mentioned above demonstrating inhibition of radioligand binding by sulfhydryl-reducing reagents. I I Mutation of the putative glycosylation sites and deletion of the intracellular carboxy-terminal tail did not affect binding. Mutations of positively charged lysine residues presumed to electrostatically bind the negatively charged ANG II were also performed.

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