Glycinin A5A4B3 mRNA: cDNA cloning and nucleotide sequencing of a splitting storage protein subunit of soybean

Momma, Takayuki; Negoro, Takaharu; Hirano, Hisashi; Matsumoto, Akiyo; Udaka, Kyoko; Fukazawa, Chikafusa

doi:10.1111/j.1432-1033.1985.tb08951.x

Cited by 64 publications

(34 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This site exactly coincides with that of the precursor proteins of several seed proteins such as glycinin (12), concanavalin A (1), and ricin (1 1). However, it is clearly different from that for the proinsulinconverting enzyme which recognizes the site just after paired basic amino acids.…”

supporting

confidence: 56%

Proglobulin Processing Enzyme in Vacuoles Isolated from Developing Pumpkin Cotyledons

Hara‐Nishimura

Nishimura

1987

Plant Physiol.

View full text Add to dashboard Cite

The enzymic conversion of proglobulin to globulin catalyzed by the extracts of vacuoles isolated from developing pumpkin (Cucurbita sp. cv Kurokawa Amakuri Nankin) cotyledons was investigated. The endoplasmic reticulum fraction isolated from the developing cotyledons pulselabeled with I35Slmethionine was shown to contain mainly the radiolabeled proglobulin, which was used as a substrate for assaying the proteolytic processing in vitro. The vacuolar extracts catalyzed the proteolytic processing of the proglobulin molecule to produce globulin containing two kinds of polypeptide chains, 'y and 5. The pH optimum for the vacuole-mediated conversion was at pH 5.0. The proteolytic processing of proglobulin by the vacuolar extracts was inhibited in the presence of various thiol reagents, e.g. p-chloromercuribenzoate, N-ethylmaleimide, iodoacetic acid, Hg2", and Cu2", but not phenylmethylsulfonyl fluoride, EDTA, o-phenanthroline, leupeptin, antipain, pepstatin, chymostatin, or pumpkin trypsin inhibitor, and was activated in the presence of dithiothreitol and cysteine, indicating that the processing enzyme is a thiol protease. The suborganellar fractionation of the vacuoles showed that the processing activity was localized in the matrix fraction, but not in the membrane or crystalloid fractions. During the seed development, the enzyme was shown to increase, exhibiting the maximal activity at the late developmental stage. The matrix fraction of the protein bodies isolated from the dry castor bean (Ricinus communis) exhibited the processing activity toward the pumpkin proglobulin molecules in the same manner as that by the matrix fraction of pumpkin vacuoles.ing of two distinct polypeptides, y and 6, linked by a disulfide bond is synthesized as precursor of a single polypeptide chain (7) and that the disulfide bond is introduced in precursor molecule synthesized in rER (5). Several other seed proteins such as lectins which are composed of two distinct polypeptide chains linked together by disulfide bond(s) are also known to be synthesized as a single polypeptide precursor, and both the in vivo labeling and cellular fractionation studies have clearly demonstrated that the protein bodies are the site of the endoproteolytic cleavage of the precursor molecules (15). However, the mechanism(s) involved in the posttranslational proteolytic processing of the single polypeptide precursor producing the mature form of protein consisting of disulfide-linked doublet polypeptide chains is not fully understood. There has been only one investigation concerning the endoproteolytic enzyme responsible for the posttranslational cleavage of the precursor proteins by Harley and Lord (9). They demonstrated that whole homogenates of the developing castor bean (Ricinus communis) endosperm contained the enzymic activities capable of converting the precursor peptides of ricin and R. communis agglutinin to their respective mature forms.In the work reported in this communication, we have examined the nature of the endoproteolytic enzyme activit...

show abstract

supporting

confidence: 56%

Proglobulin Processing Enzyme in Vacuoles Isolated from Developing Pumpkin Cotyledons

Hara‐Nishimura

Nishimura

1987

Plant Physiol.

View full text Add to dashboard Cite

show abstract

“…The susceptibility of both types of the sites mentioned above has already been observed in vitro as well as in vivo (Table 3) during the molecular maturation of 1 IS and 7 s globulin precursor polypeptides [2,[23][24][25][26][27] (and Jung et al, unpublished results). Circumstantial evidence from the literature supports this conclusion (see reviews IS, 10, 12, 281).…”

Section: Lqeeee--qshsqr---------------keeeeeeq-----------------------mentioning

confidence: 80%

Limited Proteolysis of β‐Conglycinin and Glycinin, the 7S and 11S Storage Globulins from Soybean [Glycine Max (L.) Merr.]

Shutov

Kakhovskaya

Bastrygina

et al. 1996

European Journal of Biochemistry

View full text Add to dashboard Cite

The G2 (A2BIa) glycinin subunit from soybean (Clycine mux L. Merr.) was purified and renatured to the homohexameric holoprotein. This protein along with purified P-conglycinin were subjected to limited proteolysis by trypsin. The generated polypeptide fragments were separated via SDS/PAGE and the amino acid sequence of the N-terminals was determined. Four cleavage points were detected in the a-chain A2 of glycinin as well as in the a'-chain of P-conglycinin. From the known three-dimensional structure of 7 s globulin and the hypothetical model of 7 s globulin-like 11s globulin structure, it was possible to draw the conclusion that two distinct types of susceptible sites for proteolytic cleavage are characteristic of the subunits of both globulins. The first includes the sequences linking N-and C-terminal domains of both globulins and the sequence of N-terminal extensions of 70-kDa subunits from the vicilinlike 7 s globulins. The second type includes the loop between P-strands E and F of the N-terminal domain of 11s globulins and of the C-terminal domain of 7 s globulins. A statistically significant similarity was found between the N-terminal extension of the a'-chain of P-conglycinin and the interdomain linker regions of soybean glycinin and pea legumin. It is proposed that the three sequence regions which form the first type of susceptible sites are of similar structural function and might have evolved from the Nterminal segment of a putative single-domain ancestor.Keywords: protein structure ; seed storage globulins ; gene evolution ; limited proteolysis ; Glycine rnax (L.) Merr.Glycinin and P-conglycinin from soybean seeds belong to the legumin-like 11s and vicilin-like 7s storage globulin families, respectively. Secondary structure predictions and multiple alignment of amino acid sequences [I] suggest that structural similarities should exist between 1 IS and 7s seed storage globulins. The trimer which is formed from 11s globulin subunit precursors before their cleavage into a-and /)-chains [2, 31 as well as the trimeric half-molecule from the mature hexameric 11s globulin holoprotein [4] appear to be structurally similar to the trimeric 7 s globulin molecule. X-ray-crystallographic analysis of two 7 s globulins, canavalin from jack bean [5] and phaseolin from garden bean [6, 71, revealed that their subunits are composed of two similar domains. Each domain is composed of a /]-barrel followed by a region of a-helices (Fig. 1, A1 and BI).A canonical model for the domain structure of 7 s globulins, including both 50-kDa and 70-kDa subunits, was postulated 171. However, the structural role of the extended N-terminal sequence upstream from the N-terminal domain of the 70-kDa subunits remains unclear. Different alignment methods that have been employed to compare the domains of the 7 s globulin subunits with corresponding regions from the 11s globulin subunits suggest that similar N-and C-terminal domains should exist in the legumin-like storage globulins [7, 81 (Fig. 1, A2 has been predicted that all four stru...

show abstract

“…Hypocholesterolemic activity was determined by assaying the inhibition of 3-hydroxcy-3-methylglutaryl CoA reductase (HMG-CoA reductase) in vitro and by determining the serum cholesterol levels in mice. The sequence (LPYP) was very similar to Yoshigawa's peptide (LPYPR) (Yoshikawa et al, 1999) also isolated from glycinin hydrolyzate, and the sequences including SPYPR correspond to the A 5 A 4 B 3 and A 3 B 4 regions of glycinin Momma et al, 1985). LPYPR was a little better HMG CoA reductase inhibitor than LPYP in vitro, but both reduced serum cholesterol levels by the same amount (30%) .…”

Section: Bioactive Peptidesmentioning

confidence: 73%

Fermented Soybean Products and Their Bioactive Compounds

Yang¹,

Park²,

Pak³

et al. 2011

Soybean and Health

View full text Add to dashboard Cite

Glycinin A₅A₄B₃ mRNA: cDNA cloning and nucleotide sequencing of a splitting storage protein subunit of soybean

Cited by 64 publications

References 29 publications

Proglobulin Processing Enzyme in Vacuoles Isolated from Developing Pumpkin Cotyledons

Proglobulin Processing Enzyme in Vacuoles Isolated from Developing Pumpkin Cotyledons

Limited Proteolysis of β‐Conglycinin and Glycinin, the 7S and 11S Storage Globulins from Soybean [Glycine Max (L.) Merr.]

Fermented Soybean Products and Their Bioactive Compounds

Contact Info

Product

Resources

About