Synthesis of thylakoid membrane proteins by chloroplasts isolated from spinach: Cytochrome b559 and p700-chlorophyll a-protein

Raymond E. Zielinski, C. A. Price

Research output: Contribution to journalArticle

Abstract

Intact chloroplasts, purified from spinach leaves by sedimentation in density gradients of colloidal silica, incorporate labeled amino acids into at least 16 different polypeptides of the thylakoid membranes, using light as the only source of energy. The thylakoid products of chloroplast translation were visualized by subjecting membranes purified from chloroplasts labeled with [88S]methionine to electrophoresis in high-resolution, SDS-containing acrylamide gradient slab gels and autoradiography. The apparent mol wt of the labeled products ranged from ≤10,000 to >70,000. One of the labeled products is the apoprotein of the P700-chlorophyll a-protein (CPI). The CPI apoprotein is assembled into a pigmentprotein complex which is electrophoretically indistinguishable from the native CPI complex. Isolated spinach chloroplasts also incorporate [3H]leucine and [ 35S]methionine into cytochrome b559. The radioactive label remains with the cytochrome through all stages of purification : extraction of the thylakoid membranes with Triton X-100 and urea, adsorption of impurities on DEAE cellulose, two cycles of electrophoresis in Triton-containing polyacrylamide gels and electrophoresis in SDS-containing gradient gels. Cytochrome b559 becomes labeled with both [3H]leucine and [35S]methionine and accounts for somewhat >1% of the total isotopic incorporation into thylakoid protein. The lipoprotein appears to be fully assembled during the time-course of our labeling experiments.

Original languageEnglish (US)
Pages (from-to)435
Number of pages1
JournalJournal of Cell Biology
Volume85
Issue number2
DOIs
StatePublished - May 1 1980

Fingerprint

Thylakoid Membrane Proteins
Chlorophyll Binding Proteins
Thylakoids
Spinacia oleracea
Chloroplasts
Methionine
Apoproteins
Leucine
Electrophoresis
Gels
DEAE-Cellulose
Acrylamide
Octoxynol
Cytochromes
Autoradiography
Silicon Dioxide
Lipoproteins
Adsorption
Urea
Polyacrylamide Gel Electrophoresis

ASJC Scopus subject areas

  • Cell Biology

Cite this

Synthesis of thylakoid membrane proteins by chloroplasts isolated from spinach : Cytochrome b559 and p700-chlorophyll a-protein. / Zielinski, Raymond E.; Price, C. A.

In: Journal of Cell Biology, Vol. 85, No. 2, 01.05.1980, p. 435.

Research output: Contribution to journalArticle

@article{cb974933a84b47cda30e04f7a272a93f,
title = "Synthesis of thylakoid membrane proteins by chloroplasts isolated from spinach: Cytochrome b559 and p700-chlorophyll a-protein",
abstract = "Intact chloroplasts, purified from spinach leaves by sedimentation in density gradients of colloidal silica, incorporate labeled amino acids into at least 16 different polypeptides of the thylakoid membranes, using light as the only source of energy. The thylakoid products of chloroplast translation were visualized by subjecting membranes purified from chloroplasts labeled with [88S]methionine to electrophoresis in high-resolution, SDS-containing acrylamide gradient slab gels and autoradiography. The apparent mol wt of the labeled products ranged from ≤10,000 to >70,000. One of the labeled products is the apoprotein of the P700-chlorophyll a-protein (CPI). The CPI apoprotein is assembled into a pigmentprotein complex which is electrophoretically indistinguishable from the native CPI complex. Isolated spinach chloroplasts also incorporate [3H]leucine and [ 35S]methionine into cytochrome b559. The radioactive label remains with the cytochrome through all stages of purification : extraction of the thylakoid membranes with Triton X-100 and urea, adsorption of impurities on DEAE cellulose, two cycles of electrophoresis in Triton-containing polyacrylamide gels and electrophoresis in SDS-containing gradient gels. Cytochrome b559 becomes labeled with both [3H]leucine and [35S]methionine and accounts for somewhat >1{\%} of the total isotopic incorporation into thylakoid protein. The lipoprotein appears to be fully assembled during the time-course of our labeling experiments.",
author = "Zielinski, {Raymond E.} and Price, {C. A.}",
year = "1980",
month = "5",
day = "1",
doi = "10.1083/jcb.85.2.435",
language = "English (US)",
volume = "85",
pages = "435",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "2",

}

TY - JOUR

T1 - Synthesis of thylakoid membrane proteins by chloroplasts isolated from spinach

T2 - Cytochrome b559 and p700-chlorophyll a-protein

AU - Zielinski, Raymond E.

AU - Price, C. A.

PY - 1980/5/1

Y1 - 1980/5/1

N2 - Intact chloroplasts, purified from spinach leaves by sedimentation in density gradients of colloidal silica, incorporate labeled amino acids into at least 16 different polypeptides of the thylakoid membranes, using light as the only source of energy. The thylakoid products of chloroplast translation were visualized by subjecting membranes purified from chloroplasts labeled with [88S]methionine to electrophoresis in high-resolution, SDS-containing acrylamide gradient slab gels and autoradiography. The apparent mol wt of the labeled products ranged from ≤10,000 to >70,000. One of the labeled products is the apoprotein of the P700-chlorophyll a-protein (CPI). The CPI apoprotein is assembled into a pigmentprotein complex which is electrophoretically indistinguishable from the native CPI complex. Isolated spinach chloroplasts also incorporate [3H]leucine and [ 35S]methionine into cytochrome b559. The radioactive label remains with the cytochrome through all stages of purification : extraction of the thylakoid membranes with Triton X-100 and urea, adsorption of impurities on DEAE cellulose, two cycles of electrophoresis in Triton-containing polyacrylamide gels and electrophoresis in SDS-containing gradient gels. Cytochrome b559 becomes labeled with both [3H]leucine and [35S]methionine and accounts for somewhat >1% of the total isotopic incorporation into thylakoid protein. The lipoprotein appears to be fully assembled during the time-course of our labeling experiments.

AB - Intact chloroplasts, purified from spinach leaves by sedimentation in density gradients of colloidal silica, incorporate labeled amino acids into at least 16 different polypeptides of the thylakoid membranes, using light as the only source of energy. The thylakoid products of chloroplast translation were visualized by subjecting membranes purified from chloroplasts labeled with [88S]methionine to electrophoresis in high-resolution, SDS-containing acrylamide gradient slab gels and autoradiography. The apparent mol wt of the labeled products ranged from ≤10,000 to >70,000. One of the labeled products is the apoprotein of the P700-chlorophyll a-protein (CPI). The CPI apoprotein is assembled into a pigmentprotein complex which is electrophoretically indistinguishable from the native CPI complex. Isolated spinach chloroplasts also incorporate [3H]leucine and [ 35S]methionine into cytochrome b559. The radioactive label remains with the cytochrome through all stages of purification : extraction of the thylakoid membranes with Triton X-100 and urea, adsorption of impurities on DEAE cellulose, two cycles of electrophoresis in Triton-containing polyacrylamide gels and electrophoresis in SDS-containing gradient gels. Cytochrome b559 becomes labeled with both [3H]leucine and [35S]methionine and accounts for somewhat >1% of the total isotopic incorporation into thylakoid protein. The lipoprotein appears to be fully assembled during the time-course of our labeling experiments.

UR - http://www.scopus.com/inward/record.url?scp=0019013287&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0019013287&partnerID=8YFLogxK

U2 - 10.1083/jcb.85.2.435

DO - 10.1083/jcb.85.2.435

M3 - Article

C2 - 7372715

VL - 85

SP - 435

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 2

ER -