Purification of mouse pepsinogens by pepstatin‐affinity chromatography

Esumi, Hiroyasu; Sato, Shigeaki; Sügimura, Takashi

doi:10.1016/0014-5793(78)80092-1

Cited by 18 publications

(4 citation statements)

References 13 publications

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“…The binding of full-length ProBACE (3) and ProBACE460 (shown herein) to the P10-P4Ј Stat(Val) affinity column is also consistent with the conclusion that ProBACE is catalytically active because only active forms of aspartyl proteases are expected to bind tightly to immobilized transition state analogs. Although some true zymogens of aspartyl proteases bind to immobilized transition state analogs (13)(14)(15), in these instances, the precursors invariably display markedly lower affinity for the immobilized ligand, which is not the case with ProBACE460.…”

Section: Discussionmentioning

confidence: 99%

The Pro Domain of β-Secretase Does Not Confer Strict Zymogen-like Properties but Does Assist Proper Folding of the Protease Domain

Shi¹,

Chen²,

Yin³

et al. 2001

Journal of Biological Chemistry

122

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␤-Secretase (BACE) is a membrane-bound aspartyl protease that cleaves the amyloid precursor protein to generate the N terminus of the amyloid ␤ peptide. BACE is expressed as a precursor protein containing Pre, Pro, protease, transmembrane, and cytosolic domains. A soluble BACE derivative (PreProBACE460) that is truncated between the protease and transmembrane domains was produced by baculovirus-mediated expression. ProBACE460 was purified from conditioned media of infected insect cells using immobilized concanavalin A and immobilized BACE inhibitor, P10-P4 Stat(Val). Furin cleaves ProBACE460 between the Pro and protease regions to generate mature BACE460. The k cat /K m of ProBACE460 when assayed with a polypeptide substrate is only 2.3-fold less than that of BACE460. This finding and the similar inhibitory potency of P10-P4 Stat(Val) for ProBACE460 and BACE460 suggest that the Pro domain has little effect on the BACE active site. Exposure of ProBACE460 to guanidine denaturation/ renaturation results in a 7-fold higher recovery of BACE activity than when BACE460 is similarly treated. The presence of free BACE Pro peptide during renaturation of BACE460 but not ProBACE460 increases recovery of activity. These findings show that the Pro domain in ProBACE460 does not suppress activity as in a strict zymogen but does appear to facilitate proper folding of an active protease domain.

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Section: Discussionmentioning

confidence: 99%

The Pro Domain of β-Secretase Does Not Confer Strict Zymogen-like Properties but Does Assist Proper Folding of the Protease Domain

Shi¹,

Chen²,

Yin³

et al. 2001

Journal of Biological Chemistry

122

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show abstract

“…Both type-A and type-C pepsinogens are synthesized in primates, such as humans [33] and Asian macaque monkeys [12], and in artiodactyls, such as cows [3,23] and pigs [29]. However, type-A pepsinogens are nearly the only components of pepsinogens in rabbits [19] and the Asiatic black bear [14], whereas type-C pepsinogens are the exclusive components of pepsinogens in rodents, such as mice [5], rats [9], and guinea pigs [13]. Multiple isozymogens have been found for both type-A and type-C pepsinogens [6,21].…”

Section: Introductionmentioning

confidence: 99%

Multiplicities and some enzymatic characteristics of ape pepsinogens and pepsins

Narita

Oda

Takenaka

et al. 2000

J of Medical Primatology

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Pepsinogen levels in ape stomachs were comparable to those in macaques and significantly higher than those in the stomachs of other mammals, including carnivores and ruminants. The occurrence of multiple forms of pepsinogens was remarkable. Nine, sixteen, eight, and fourteen pepsinogens were purified or partially purified from the gastric mucosa of a gibbon, orang-utan, gorilla, and chimpanzee, respectively. Most of these were type-A pepsinogens, and only one type-C pepsinogen was identified in each ape. The two types could be readily distinguished by staining for proteolytic activity on polyacrylamide gel electrophoresis (PAGE) in the presence/absence of pepstatin. Type-A pepsinogens were further divided into two subtypes. One subtype, constituting a major group of pepsinogens in apes, exhibited high hemoglobin-digestive activity. The other subtype was specified by a relatively high content of Lys and low hemoglobin-digestive activity. It is likely that pepsinogen-A genes have been duplicated several times as hominoids, including humans, evolved in the primate lineage. The presence of multiple pepsinogens in apes might be advantageous in the efficient digestion of a wide variety of foods.

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“…The molecular weight of the major band corresponding to the enzyme was estimated to be approximately 38 kDa under reducing conditions which was higher than that of the camel pepsin purified by Abuharfeel and Abuereish (1984) and Al-Mzaien (2007) and also higher than these from cattle (Fox et al, 1977), sheep (Fox et al, 1977), Japanese monkey (Kageyama and Takahashi, 1976), mouse (Esumi et al, 1978) and pigs (Rajagopalan et al, 1966). This difference could be the result of post-translational modifications such as phosphorylation.…”

Section: The Purification Of Milk Clotting Enzyme From Adult Camel Abmentioning

confidence: 93%

Molecular characterization of crude enzymatic extract from Algerian camel abomasum (Camelus dromedarius)

Souad

Saliha

Nouani

et al. 2016

Emir. J. Food Agric

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Purification of mouse pepsinogens by pepstatin‐affinity chromatography

Cited by 18 publications

References 13 publications

The Pro Domain of β-Secretase Does Not Confer Strict Zymogen-like Properties but Does Assist Proper Folding of the Protease Domain

The Pro Domain of β-Secretase Does Not Confer Strict Zymogen-like Properties but Does Assist Proper Folding of the Protease Domain

Multiplicities and some enzymatic characteristics of ape pepsinogens and pepsins

Molecular characterization of crude enzymatic extract from Algerian camel abomasum (Camelus dromedarius)

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