Biochemical and immunochemical studies of α-amylase from the salivary glands of the mouse

Amerongen, A. van Nieuw; Oderkerk, C.H.; Vreugdenhil, Angie; Roukema, P.A.

doi:10.1016/0003-9969(79)90222-x

Cited by 12 publications

(4 citation statements)

References 27 publications

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“…This is first time that two species, glycosylated and non‐glycosylated, of recombinant mouse α‐amylases were purified separately to homogeneous preparations. Native (non‐recombinant) mouse α‐amylase was also secreted in two forms, glycosylated and non‐glycosylated (Hjorth, 1979; Nieuw Amerongen, et al ., 1979). As shown in Table 1, non‐glycosylated α‐amylase (229 U/mg protein) in the final preparation showed specific activity about two‐fold higher than that of the glycosylated product (105 U/mg protein).…”

Section: Resultsmentioning

confidence: 99%

Efficient expression, purification and characterization of mouse salivary α‐amylase secreted from methylotrophic yeast, Pichia pastoris

Kato

Ishibashi

Tatsuda

et al. 2001

Yeast

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We constructed a secretion vector of mouse salivary a-amylase, pPAM, using the AOX1 promoter-terminator and the secretion signal of 128 kDa pGKL killer protein, for an alternative yeast, Pichia pastoris. Taking advantage of multicopy insertion of the expression cassette and optimized growth conditions, we succeeded in highly ef®cient extracellular production (y240 mg/ml) of mouse a-amylase in the 10 ml scale by conventional¯ask culture: this ef®ciency was about 90-fold higher than that of Saccharomyces cerevisiae. Growth temperature of cells was critical for ef®cient production of a-amylase. P. pastoris transformants secreted both core-glycosylated and nonglycosylated a-amylase molecules with a glycosylated : non-glycosylated ratio of about 20 : 80. Both glycosylated and non-glycosylated a-amylases were puri®ed separately to apparent homogeneity. The signal sequence was correctly processed in both species, and the molecular masses of glycosylated and non-glycosylated a-amylase were determined to be 58 600 and 56 300, respectively, by mass spectrometry. We further studied the outer chain glycosylation of engineered mouse a-amylase secreted by P. pastoris.

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Section: Resultsmentioning

confidence: 99%

Efficient expression, purification and characterization of mouse salivary α‐amylase secreted from methylotrophic yeast, Pichia pastoris

Kato

Ishibashi

Tatsuda

et al. 2001

Yeast

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“…Since pancreata of M 1 /M 3 receptor double KO mice contained similar amounts of amylase as pancreata of WT mice, it is likely that the lack of AChstimulated pancreatic amylase release is responsible, at least partially, for the reduction in serum amylase levels displayed by the M 1 /M 3 receptor double KO mice. Besides the exocrine pancreas, salivary glands, particularly the parotid gland, also release substantial amounts of amylase into the circulation (Amerongen et al, 1980). We and Nakamura et al (2004) recently demonstrated that muscarinic agonist (pilocarpine)-induced increases in salivary flow are abolished in M 1 /M 3 receptor double KO mice.…”

Section: Discussionmentioning

confidence: 98%

“…6). Several studies suggest that the exocrine pancreas and the salivary glands are the major sources of serum amylase (Amerongen et al, 1980;Pieper-Bigelow et al, 1990). Pancreatic amylase is predicted to enter the circulation via a direct leak from pancreatic acini or ducts into the blood (Pieper-Bigelow et al, 1990).…”

Section: Discussionmentioning

confidence: 99%

Cholinergic Stimulation of Amylase Secretion from Pancreatic Acinar Cells Studied with Muscarinic Acetylcholine Receptor Mutant Mice

Gautam¹,

Han²,

Heard³

et al. 2005

J Pharmacol Exp Ther

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Muscarinic acetylcholine receptors (mAChRs) expressed by pancreatic acinar cells play an important role in mediating acetylcholine-dependent stimulation of digestive enzyme secretion. To examine the potential roles of M 1 and M 3 mAChRs in this activity, we used M 1 and M 3 receptor single knockout (KO) and M 1 /M 3 receptor double KO mice as novel experimental tools. Specifically, we examined the ability of the muscarinic agonist carbachol to stimulate amylase secretion in vitro, using dispersed pancreatic acini prepared from wild-type and mAChR mutant mice. Quantitative reverse transcription-polymerase chain reaction studies using RNA prepared from mouse pancreatic acini showed that deletion of the M 1 or M 3 mAChR genes did not lead to significantly altered mRNA levels of the remaining mAChR subtypes. Moreover, immunoprecipitation studies with M 1 and M 3 mAChR-selective antisera demonstrated that both mAChR subtypes are expressed by mouse pancreatic acini. Strikingly, carbachol-induced stimulation of amylase secretion was significantly impaired in acinar preparations from both M 1 and M 3 receptor single KO mice and abolished in acinar preparations from M 1 /M 3 receptor double KO mice. However, another pancreatic secretagogue, bombesin, retained its ability to fully stimulate amylase secretion in acinar preparations from M 1 /M 3 receptor double KO mice. Together, these studies support the concept that cholinergic stimulation of pancreatic amylase secretion is mediated by a mixture of M 1 and M 3 mAChRs and that other mAChR subtypes do not make a significant contribution to this activity. These findings clarify the long-standing question regarding the molecular nature of the mAChR subtypes mediating the secretion of digestive enzymes from the exocrine pancreas.

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“…We think that the source organ of these serum amylases in mastomys may be the parotid gland. Isoamylases in the saliva, parotid gland and pancreas of mastomys were detected in these organs of mouse (Nieuw Amerongen et al, 1979;Ikeno et al, 1988) and rat (Ikeno et al, 1982;Murata et al, 1986).…”

Section: Resultsmentioning

confidence: 99%

An electrophoretic polymorphism in salivary amylases (Amy-1) of mastomys (Praomys coucha)

Matsushima

Ikeno

et al. 1990

Lab Anim

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SummaryAn electrophoretic polymorphism of salivary amylases (Amy-I) in mastomys (Praomys coucha) (MWC, MRJ and MCC strains) was detected. Amylase in MWC or MRJ saliva, which migrated fast toward the anode, was designated as AMY-lA, and that in MCC saliva migrating slowly as AMY -lB. Salivary amylases are controlled by a pair of codominant alleles at a single autosomal locus (Amy-I). No polymorphism was seen in pancreatic amylases . The frequencies of these phenotypes did not differ between the sexes. Some isoamylases were observed and these were different from those in mouse or rat.

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Biochemical and immunochemical studies of α-amylase from the salivary glands of the mouse

Cited by 12 publications

References 27 publications

Efficient expression, purification and characterization of mouse salivary α‐amylase secreted from methylotrophic yeast, Pichia pastoris

Efficient expression, purification and characterization of mouse salivary α‐amylase secreted from methylotrophic yeast, Pichia pastoris

Cholinergic Stimulation of Amylase Secretion from Pancreatic Acinar Cells Studied with Muscarinic Acetylcholine Receptor Mutant Mice

An electrophoretic polymorphism in salivary amylases (Amy-1) of mastomys (Praomys coucha)

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