Furin, PC1/3, and/or PC6A Process Rabbit, but Not Human, Pro-lactase-phlorizin Hydrolase to the 180-kDa Intermediate

Keller, Patrick; Zecca, Laura; Boukamel, Rita; Zwicker, Elmar; Gloor, Sergio M.; Semenza, Giorgio

doi:10.1074/jbc.270.43.25722

Cited by 9 publications

(9 citation statements)

References 58 publications

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“…Although number, types, and cellular sites of the proteolytic events producing mature LPH from prepro-LPH have been studied extensively, albeit with controversial results, much less has been learned about the proteases implicated in the maturation. A central question in our investigation was whether PCs of the furin type may partake in the series of events forming mature LPH from human pro-LPH, as we have shown for the 180-kDa intermediate occurring in the rabbit (23).…”

Section: Discussionmentioning

confidence: 99%

“…Expressing Furin-We have shown previously that predominantly furin, PC1/3, and/or PC6A are implicated in the processing of rabbit pro-LPH to generate the 180-kDa intermediate form (23). We therefore investigated whether furin generates human mature-like LPH.…”

Section: Generation Of Mature-like Lph Form In Cos-7 Cellsmentioning

confidence: 99%

“…To study the processing of human prepro-LPH in detail we used COS-7 cells as a model. This cell line was used successfully in our laboratory to study the processing of rabbit lactase (23). Fig.…”

Section: Processing Of Human Prepro-lph and Pro-lph In Trans-mentioning

confidence: 99%

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Processing of Human Intestinal Prolactase to an Intermediate Form by Furin or by a Furin-like Proprotein Convertase

Mesonero

Gloor

Semenza³

1998

Journal of Biological Chemistry

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Human lactase-phlorizin hydrolase (human-LPH) is synthesized as a large precursor (prepro-LPH), then cleaved to a pro-LPH of 220 kDa which is further cut to a "mature-like LPH" of a size close to that of mature LPH, i.e. about 150 kDa (in the processing of rabbit pro-LPH the intermediate has a mass of approximately 180 kDa). By coexpression of human prepro-LPH with furin in COS-7 cells we show that furin generates a mature-like LPH. Radioactive amino acid sequence analysis reveals that furin recognizes the motif R-T-P-R 832 , a protein convertase consensus, to generate a NH 2 terminus located 36 amino acids upstream of the NH 2 terminal found in vivo at Ala 869 . This intermediate is ultimately cleaved to the mature LPH form by other proteases including the pancreatic ones. These data demonstrate that human pro-LPH, like the rabbit enzyme, is processed to the mature enzyme by furin or furin-like enzymes through at least an intermediate form that has, however, an apparent mass close to that of the mature enzyme.

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Section: Discussionmentioning

confidence: 99%

Section: Generation Of Mature-like Lph Form In Cos-7 Cellsmentioning

confidence: 99%

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Processing of Human Intestinal Prolactase to an Intermediate Form by Furin or by a Furin-like Proprotein Convertase

Mesonero

Gloor

Semenza³

1998

Journal of Biological Chemistry

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“…Furthermore the tandem association of domains I and II is required for the LPHα function, suggestive of a role in folding or in the acquisition of the quaternary structure of the mature enzyme. In rat and rabbit, unlike human, LPHα is subject to internal cleavage during precursor maturation [4,5,29,30]. The functional role (if any) of this cleavage is unknown, but might be related to the different pattern of lactase expression observed along the villus axis in human compared with rat and rabbit ; indeed some enterocytes in hypolactasic humans contain LPH mRNA but no protein, whereas other enterocytes express an inactive protein [55][56][57].…”

Section: Role Of Lphαmentioning

confidence: 99%

“…In rat and rabbit, in contrast with human, internal cleavage by a membrane protease of the furin family occurs within LPHα [4,29,30]. Studies on the human precursor suggested an obligate requirement of LPHα for the production of enzymically active and transport-competent LPHβ [28,31].…”

Section: Introductionmentioning

confidence: 99%

Functional diversity and interactions between the repeat domains of rat intestinal lactase

Jost

Duluc

Richardson

et al. 1997

Biochemical Journal

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Lactase-phlorizin hydrolase (LPH), a major digestive enzyme in the small intestine of newborns, is synthesized as a highmolecular-mass precursor comprising four tandemly repeated domains. Proteolytic cleavage of the precursor liberates the pro segment (LPHα) corresponding to domains I and II and devoid of known enzymic function. The mature enzyme (LPHβ) comprises domains III and IV and is anchored in the brush border membrane via a C-terminal hydrophobic segment. To analyse the roles of the different domains of LPHα and LPHβ, and the interactions between them, we have engineered a series of modified derivatives of the rat LPH precursor. These were expressed in cultured cells under the control of a cytomegalovirus promoter. The results show that recombinant LPHβ harbouring both domains III and IV produces lactase activity. Neither domain III nor IV is alone sufficient to generate active enzyme, although the corresponding proteins are transport-competent.

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In vitro translation of RNA to lactase during postnatal development of rat intestine

Kaur

Mahmood

et al. 2005

J. Biosci.

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mRNA levels encoding lactase were detected by Northern blot analysis using two different probes in developing rat intestine. Probe I and probe II corresponding to second half of prolactase gene showed a 6.8 kb mRNA transcript in 7, 14, 21 and 30 day old rat intestine. There was no change in quantity of lactase mRNA detected using probe II, but hybridization with probe I showed a progressive decrease in mRNA transcript encoding lactase with age. At day 7 and 14 of postnatal development, the lactase mRNA was quite high, but it reduced upon weaning. The in vitro translation products of RNA detected by Western blot analysis using brush border lactase antibodies showed several isoforms of lactase antigen with molecular weight ranging from 100-220 kDa. Analysed at days 7 and 30 of postnatal development, lactase isoforms of molecular weight 130 kDa and 220 kDa were similar to those found in purified brush border membranes. The translation of RNA to 220 kDa lactase protein was high in 7 and 14 day old pups, but it was markedly reduced in 30 day old animals. These findings support the contention that translation of mRNA to lactase is impaired in weaned animals, which may also be responsible for the maturational decline in lactase activity in adult rat intestine.

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Furin, PC1/3, and/or PC6A Process Rabbit, but Not Human, Pro-lactase-phlorizin Hydrolase to the 180-kDa Intermediate

Cited by 9 publications

References 58 publications

Processing of Human Intestinal Prolactase to an Intermediate Form by Furin or by a Furin-like Proprotein Convertase

Processing of Human Intestinal Prolactase to an Intermediate Form by Furin or by a Furin-like Proprotein Convertase

Functional diversity and interactions between the repeat domains of rat intestinal lactase

In vitro translation of RNA to lactase during postnatal development of rat intestine

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