Parathyroid hormone (PTH) and guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma S) increase levels of the second messenger inositol 1,4,5-triphosphate (IP3) and other inositol phosphates (IP) in several membrane preparations of PTH-responsive cells. We present evidence here indicating that in a membrane preparation of canine renal cortical tubular cells bPTH-(1-84), bPTH-(1-34), [N-Leu8,18Tyr34]bPTH-(3-34)NH2, and the human PTH related peptide fragment hPTHrP-(1-34)NH2 all increase levels of inositol phosphate (IP) but [Tyr34]-bPTH-(7-34)NH2 and hPTHrP-(7-34)NH2 have no significant effects on IP accumulation. Increases in IPs are generally attributed to increased formation of IPs and appear to be mediated by a G protein. However, increased levels of IPs may also result from inhibition of the phosphatases are responsible for their metabolism. We investigated the effect of PTH and GTP-gamma S on the metabolism of IP3 in canine renal cortical tubular membranes. These membranes rapidly metabolize [3H]IP3 (47% at 15 s). Decreases in [3H]IP3 at all time points are accounted for quantitatively by increases in the sum of its breakdown products: [3H]IP2, [3H]IP1, and [3H]inositol. After 5 minutes of exposure to membranes, the vast majority of [3H]IP3 (84%) is converted to its terminal metabolite, [3H]inositol. GTP-gamma S (100 microM) inhibits the amount of [3H]IP3 metabolized in 15 s by 70% and reduces the amount of [3H]inositol ultimately formed in 5 minutes by 64%. ATP-gamma S, ATP, and 2,3-bisphosphoglycerate (100 microM) also inhibit [3H]IP3 hydrolysis in this preparation.(ABSTRACT TRUNCATED AT 250 WORDS)