Synthesis and partial maturation of the α‐ and γ‐subunits of the mouse submaxillary gland nerve growth factor in <i>Xenopus laevis</i> oocytes

Wion, Didier; Dicou, Eleni; Brachet, Philippe

doi:10.1016/0014-5793(84)80053-8

Cited by 11 publications

(9 citation statements)

References 23 publications

(9 reference statements)

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“…The NGF transcript found in the thyroid is identical in size to that of the MSG and to those characterized in other peripheral tissues (7,8).…”

Section: Discussionmentioning

confidence: 63%

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Synthesis of nerve growth factor mRNA and precursor protein in the thyroid and parathyroid glands of the rat.

Dicou¹,

Lee²,

Brachet³

1986

Proc. Natl. Acad. Sci. U.S.A.

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Nerve growth factor (NGF) mRNAs were detected in the thyroid and parathyroid glands by hybridization to a preproNGF and a NGF-,/ cDNA probe. The thyroid NGF transcript was identical in size to that found in the mouse submaxillary gland. Mature NGF-13 was not detected in the thyroid tissue either by immunoprecipitation or by immunohistochemical methods. In contrast, an antiserum directed against the predicted precursor protein sequence immunoprecipitated a protein of Mr 30,000-31,000 in thyroid extracts. A protein of identical molecular weight was also detected by this serum in the mouse submaxillary gland extracts. In vitro translation of hybrid-selected NGF RNAs detected two peptides ofMrs 35,000 and 28,000. These peptides closely correspond to the proposed translation products of the preproNGF mRNA sequence, suggesting that two different initiation codons can be utilized, at least in vitro. The proNGF antiserum also cross-reacted with the in vitro synthesized Mr 35,000 peptide. The antigenic determinant recognized by the precursor-specific antibody was localized to the convoluted granular tubules of the mouse submaxillary gland, known to be the site of synthesis of the NGF-,8. We present evidence for the localization of unprocessed proNGF in the parathyroid gland and in some parafollicular cells of the thyroid.Type ,3 nerve growth factor (NGF-13) is a trophic factor that plays a key role in the development and survival of sympathetic and sensory neurons (1). It is synthesized at exceptionally high levels in the male mouse submaxillary gland (MSG) (2), where it associates with other protein subunits to form a 7S complex (3).The amino acid sequence of the mouse NGF-/3 has been determined (4), and the cDNA encoding its mRNA has been isolated (5, 6). The nucleotide sequence of the cDNA clone predicts that NGF-P3 is synthesized as part of a larger precursor, preproNGF. This molecule may be cleaved at dibasic residues to generate three additional peptides besides the NGF-,3 peptide that is situated at the carboxyl terminus.However, the identification of the in vivo NGF forms has been a difficult task. This was mainly due to the fact that NGF-,B antisera show no specificity for the NGF precursor protein(s) (7,8).

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“…The NGF transcript found in the thyroid is identical in size to that of the MSG and to those characterized in other peripheral tissues (7,8).…”

Section: Discussionmentioning

confidence: 63%

“…This was mainly due to the fact that NGF-,B antisera show no specificity for the NGF precursor protein(s) (7,8). …”

Section: Introductionmentioning

confidence: 99%

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Synthesis of nerve growth factor mRNA and precursor protein in the thyroid and parathyroid glands of the rat.

Dicou¹,

Lee²,

Brachet³

1986

Proc. Natl. Acad. Sci. U.S.A.

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“…That mRNA NGF levels precede the corresponding increases in NGF levels for several days is unique to the septo-hippocampal system, and it has to be assumed that either the mRNA ~GF is initially present in a nontranslatable form, or that, in the hippocampus during the period of the most rapid m R N A NGF increase, the m R N A NGF level is not rate-limiting for the production of NGE If for instance the proteolytic processing enzymes for the NGF precursor were rate-limiting, a build-up of NGF precursor molecules would take place. These molecules are not recognized by antibodies directed against mature NGF and they also seem to be biologically inactive (Wion et al 1984;Dicou et al 1986). In addition to the unusually long time-lag between the increase in mRNA NG~ and the subsequent increase in NGF levels, the time-course of the perinatal development of hippocampal NGF and mRNA NGF showed additional unexpected features.…”

Section: Comparison Between Developmental Changes Of Ngf and Chat In mentioning

confidence: 99%

The physiological function of nerve growth factor in the central nervous system: Comparison with the periphery

Thoenen¹,

Bandtlow²,

Heumann³

1987

Reviews of Physiology, Biochemistry and Pharmacology

507

175

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“…Our antisera to NGF do not recognize the NGF precursor [4,5]. Therefore, in order to biochemically characterize this protein and to identify cell types that synthesize it, our strategy consisted of raising antibodies to synthetic peptides that reproduce sequences of this molecule.…”

Section: Introductionmentioning

confidence: 99%

Interaction of antibodies to synthetic peptides of proNGF with in vitro synthesized NGF precursors

Dicou

1989

FEBS Letters

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Sera raised against three synthetic peptides that reproduce sequences of the pro-nerve powth factor (proNGF) protein were tested in immunoprecipitation experiments using in vitro translation products of SP6-directed NGF mRNA in a rabbit reticulocyte lysate. The interaction of these antibodies with bacterially synthesixed chimeric preproNGF was also examined. Digestion of the translation products by the y-subunit generated the 22 and 18 kDa intermediates. A predominant 13 kDa intermediate was obtained after digestion of translation products in wheat germ extract. This is shown to be the N-terminal peptide by immunoprecipitation with an anti-peptide serum. These antibodies may be used to detect NGF precursor cleavage products in vivo.

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Synthesis and partial maturation of the α‐ and γ‐subunits of the mouse submaxillary gland nerve growth factor in Xenopus laevis oocytes

Cited by 11 publications

References 23 publications

Synthesis of nerve growth factor mRNA and precursor protein in the thyroid and parathyroid glands of the rat.

Synthesis of nerve growth factor mRNA and precursor protein in the thyroid and parathyroid glands of the rat.

The physiological function of nerve growth factor in the central nervous system: Comparison with the periphery

Interaction of antibodies to synthetic peptides of proNGF with in vitro synthesized NGF precursors

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