Identification of glycinin in vivo as a polyamine-conjugated protein via a γ-glutamyl linkage

Hyeog Kang, Seung Kwan Lee, Young Dong Cho

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

To identify a polyamine-conjugated protein by the action of transglutaminase in the absence of radiolabelled polyamine, extracts prepared from the leaves and developing soybean seeds were investigated for the specific activity of transglutaminase and the content of free polyamines. We identified the major storage protein, glycinin, as a polyamine-conjugated protein. This was established by the following procedures: (1) immunolocalization with antibody against putrescine prepared in rabbit against putrescine-BSA conjugate; (2) immunocross-reactivity on nitrocellulose transblot of the purified glycinin subunits by using antibody against putrescine; (3) identification of polyamines in acid hydrolysates of purified glycinin; (4) release of polyamines in proteolytic digests through the catalytic action of γ-glutamylamine cyclotransferase, an enzyme specific for the disassembly of γ-glutamylamines. The activity of γ-glutamylamine cyclo-transferase was also identified in soybean seeds.

Original languageEnglish
Pages (from-to)467-473
Number of pages7
JournalBiochemical Journal
Volume332
Issue number2
DOIs
Publication statusPublished - 1998 Jun 1
Externally publishedYes

Fingerprint

Polyamines
Putrescine
Proteins
Transglutaminases
Soybeans
Seed
Seeds
Collodion
Antibodies
Transferases
glycinin
Rabbits
Acids
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Identification of glycinin in vivo as a polyamine-conjugated protein via a γ-glutamyl linkage. / Kang, Hyeog; Lee, Seung Kwan; Cho, Young Dong.

In: Biochemical Journal, Vol. 332, No. 2, 01.06.1998, p. 467-473.

Research output: Contribution to journalArticle

@article{ef017f866b064cc89a9ab4e142e98c37,
title = "Identification of glycinin in vivo as a polyamine-conjugated protein via a γ-glutamyl linkage",
abstract = "To identify a polyamine-conjugated protein by the action of transglutaminase in the absence of radiolabelled polyamine, extracts prepared from the leaves and developing soybean seeds were investigated for the specific activity of transglutaminase and the content of free polyamines. We identified the major storage protein, glycinin, as a polyamine-conjugated protein. This was established by the following procedures: (1) immunolocalization with antibody against putrescine prepared in rabbit against putrescine-BSA conjugate; (2) immunocross-reactivity on nitrocellulose transblot of the purified glycinin subunits by using antibody against putrescine; (3) identification of polyamines in acid hydrolysates of purified glycinin; (4) release of polyamines in proteolytic digests through the catalytic action of γ-glutamylamine cyclotransferase, an enzyme specific for the disassembly of γ-glutamylamines. The activity of γ-glutamylamine cyclo-transferase was also identified in soybean seeds.",
author = "Hyeog Kang and Lee, {Seung Kwan} and Cho, {Young Dong}",
year = "1998",
month = "6",
day = "1",
doi = "10.1042/bj3320467",
language = "English",
volume = "332",
pages = "467--473",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - Identification of glycinin in vivo as a polyamine-conjugated protein via a γ-glutamyl linkage

AU - Kang, Hyeog

AU - Lee, Seung Kwan

AU - Cho, Young Dong

PY - 1998/6/1

Y1 - 1998/6/1

N2 - To identify a polyamine-conjugated protein by the action of transglutaminase in the absence of radiolabelled polyamine, extracts prepared from the leaves and developing soybean seeds were investigated for the specific activity of transglutaminase and the content of free polyamines. We identified the major storage protein, glycinin, as a polyamine-conjugated protein. This was established by the following procedures: (1) immunolocalization with antibody against putrescine prepared in rabbit against putrescine-BSA conjugate; (2) immunocross-reactivity on nitrocellulose transblot of the purified glycinin subunits by using antibody against putrescine; (3) identification of polyamines in acid hydrolysates of purified glycinin; (4) release of polyamines in proteolytic digests through the catalytic action of γ-glutamylamine cyclotransferase, an enzyme specific for the disassembly of γ-glutamylamines. The activity of γ-glutamylamine cyclo-transferase was also identified in soybean seeds.

AB - To identify a polyamine-conjugated protein by the action of transglutaminase in the absence of radiolabelled polyamine, extracts prepared from the leaves and developing soybean seeds were investigated for the specific activity of transglutaminase and the content of free polyamines. We identified the major storage protein, glycinin, as a polyamine-conjugated protein. This was established by the following procedures: (1) immunolocalization with antibody against putrescine prepared in rabbit against putrescine-BSA conjugate; (2) immunocross-reactivity on nitrocellulose transblot of the purified glycinin subunits by using antibody against putrescine; (3) identification of polyamines in acid hydrolysates of purified glycinin; (4) release of polyamines in proteolytic digests through the catalytic action of γ-glutamylamine cyclotransferase, an enzyme specific for the disassembly of γ-glutamylamines. The activity of γ-glutamylamine cyclo-transferase was also identified in soybean seeds.

UR - http://www.scopus.com/inward/record.url?scp=0032102524&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032102524&partnerID=8YFLogxK

U2 - 10.1042/bj3320467

DO - 10.1042/bj3320467

M3 - Article

C2 - 9601076

AN - SCOPUS:0032102524

VL - 332

SP - 467

EP - 473

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -