Microscopical localization of adenylate cyclase: A historical review of methodologies

Product identification and adenylyl cyclase activity in chloroplasts of Nicotiana tabacum

“…Sampling and extraction techniques were altered to minimise physicochemical and biological contamination of the authentic plant derived cyclic nucleotide measurements. Additionally, alternative triphosphonucleotide substrates, like adenylyl diphosphoimidophosphate or adenylyl diphosphoimidomethyl phosphate, were used as adenylyl cyclase specific substrates 4,5. Furthermore, the endogenous adenylyl cyclase activity is low and in absolute value per sample the amount of its product is often at the sub‐picomole level,6–11 so that even sensitive methods have to be applied near their detection limit.…”

“…Adenylyl cyclase activity has been demonstrated in higher plants by the use of histochemical and biochemical methods. Histochemical studies contribute greatly in revealing topological information on AC activity at the ultra‐structural level and its association with specific tissues and organelles, and help find clues to its physiological function 5. The only molecular biological evidence is a recently isolated cDNA encoding for a soluble signalling protein with AC activity 17.…”

Tansley Review No. 106

“…The specificity of the reaction was considerably improved by the replacement of ATP with a specific substrate analogue for adenylyl cyclase such as adenosine 5h- [β,γ-imido] triphosphate (Yount et al, 1971) or adenosine 5h-[α,β-methylene]triphosphate (Mayer et al, 1985), which are much less sensitive to ATPase and other phosphatase activities. (For a recent review on histochemical methodology the reader is referred to Richards & Richards (1998).) By such methods, early indications of the presence of adenylyl cyclase activity were found in plasma membrane, in endoplasmic reticulum and nuclear membranes in Zea mays root tips (Al-azzawi & Hall, 1976), on internal membranes of cytoplasmatic vacuoles in Pisum sativum (Hilton & Nesius, 1978), on the external side of the host plasma membrane and membranes surrounding the endophyte in root nodules of Alnus glutinosa (Gardner et al, 1979), and on the external side of the plasma membrane of Pisum sativum (Nougare ' de et al, 1984).…”