Phosphate transport in yeast mitochondria: purification and characterization of a mitoribosomal synthesis dependent proteolipid showing a high affinity for phosphate

Guérin, Martine; Napias, Christian

doi:10.1021/bi00606a009

Cited by 44 publications

(21 citation statements)

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“…For example, carrier proteins for phosphate, chloride, and sulfate transport have been recently identified and isolated from bacteria (1,20), fungi (6), and human red blood cells (see reviews 10,24). In the red blood cell studies (2-4, 8, 11, 17), several nonpermanent, amino-reactive reagents (e.g., SITS,2 DIDS, DADS, and NAP-taurine), which covalently bind to the plasmalemma, have been used to identify and characterize the band 3 membrane protein as the carrier for CF-and s042-transport.…”

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confidence: 99%

Inhibition of Anion Transport in Corn Root Protoplasts

Lin¹

1981

Plant Physiol.

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MATERIALS AND METHODSThe effects of several amino-reactive disulfonic stilbene deriatives and N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate on Clr, S042-, and inorganic phosphate (Pi) uptake in protoplasts isolated from corn root tissue were studied. 4-Acetamido-4'-isothiocyano-2,2'-stilbenedisulfonic acid, 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid, 4,4'-diamino-2,2'-stilbenedisulfonic acid, and NAP-taurine inhibited Cl and S042-but not Pi and K+ uptake in corn root protoplasts; whereas mersalyl inhibited Pi but not Cl1 or S04-2 uptake. The rate of uptake of all anions decreased with increasing external pH. In addition, these reagents markedly inhibited plasmalemma ATPase activity isolated from corn root tissue. Excised root segments were less sensitive to Cl and S042-transport inhibitors.A growing interest has emerged on the nature of anion transport in several biological systems. For example, carrier proteins for phosphate, chloride, and sulfate transport have been recently identified and isolated from bacteria (1, 20), fungi (6), and human red blood cells (see reviews 10, 24). In the red blood cell studies (2-4, 8, 11, 17), several nonpermanent, amino-reactive reagents (e.g., SITS,2 DIDS, DADS, and NAP-taurine), which covalently bind to the plasmalemma, have been used to identify and characterize the band 3 membrane protein as the carrier for CF-and s042-transport.Virtually no information is available on the nature of the carrier proteins responsible for ion transport in plant cells, particularly root cells which are the major sites of entry for ions into the plants. The lack of such information is due, in part, to the lack of suitable technique for isolating functional protoplasts from roots which are, by their nature, more amenable to techniques that have been used to characterize membrane transport. Recently, we developed techniques for the large scale and rapid isolation of protoplasts from corn roots (13) which are capable of transporting several ions (K+, H2PO4 , and H+). In this study, several chemical modifiers which have been used to study anion transport into red blood cells were employed to study anion transport into corn root protoplasts with the intent of providing further insight into anion transport in plant cells. Delaware, 19801. 2Abbreviations: SITS, 4-acetamido4'-isothiocyano-2,2'-stilbenedisulfonic acid; DIDS: 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid; DADS, 4,4'-diamino-2,2'-stilbenedisulfonic acid; NAP-taurine, N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate; FCCP, (p-trifluoromethoxy) carbonyl cyanide phenylhydrazone; DIFP, diisopropylfluorophosphate.A modification of our previously reported method (13) was used to isolate protoplasts from young corn roots. As outlined in Figure 1, 0.1% pectolyase Y-23 (16) substituted for pectinase and hemicellulase in the digestion enzyme mixture. Also, the tissue incubation time in the digestion mixture was shortened to 1.5 to 2 h and the precentrifugation steps were omitted. The yield of 106 protoplasts/g root tissue and...

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confidence: 99%

Inhibition of Anion Transport in Corn Root Protoplasts

Lin¹

1981

Plant Physiol.

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“…In [28,29] it was suggested that one proteolipid could be the Pi binding proteolipid. The possibility that the 10 000 M r band was the Pi-binding proteolipid [ 12,13 ] is under investigation.…”

Section: Resultsmentioning

confidence: 99%

“…From initial observations [11 ] a proteolipid having affinity for Pi was extracted from yeast mitochondria by organic solvents [12]. This proteolipid is mitochondrially translated and its participation in phosphate transport has been suggested [12,13].…”

Section: Introductionmentioning

confidence: 99%

“…This proteolipid is mitochondrially translated and its participation in phosphate transport has been suggested [12,13]. This Pi binding proteolipid is, however, different from the DCCD-binding protein [ 13].…”

Section: Introductionmentioning

confidence: 99%

“…This Pi binding proteolipid is, however, different from the DCCD-binding protein [ 13]. However, a chloroform extract able to bind Pi was obtained directly from the ATPase complex [12].…”

Section: Introductionmentioning

confidence: 99%

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Existence of two different species of mitochondrially translated proteolipids in ATPase complex of yeast mitochondria

1981

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Amino acid sequence of a new mitochondrially synthesized proteolipid of the ATP synthase of Saccharomyces cerevisiae.

Velours¹,

Esparza²,

Hoppe³

et al. 1984

The EMBO Journal

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The purification and the amino acid sequence of a proteolipid translated on ribosomes in yeast mitochondria is reported. This protein, which is a subunit of the A TP synthase, was purified by extraction with chloroform/methanol (2/1) and subsequent chromatography on phosphocellulose and reverse phase h.p.l.c. A mol. wt. of 5500 was estimated by chromatography on Bio-Gel P-30 in 8011/o fonnie acid. The complete amino acid sequence of this protein was determined by automated solid phase Edman degradation of the whole protein and of fragments obtained after cleavage with cyanogen bromide. The sequence analysis indicates a length of 48 amino acid residues. The calculated mol. wt. of 5870 corresponds to the value found by gel chromatography. This polypeptide contains three basic residues and no negatively charged side chain. The three basic residues are clustered at the C terminus. The primary structure of this protein is in full agreement with the predicted amino acid sequence of the putative polypeptide encoded by the mitochondrial aap1 gene recently discovered in Saccharomyces cerevisiae. Moreover, this protein shows 5011/o homology with the amino acid sequence of a putative polypeptide encoded by an unidentified reading frame also discovered near the mitochondrial ATPase subunit 6 genein Aspergillus nidulans.

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Phosphate transport in yeast mitochondria: purification and characterization of a mitoribosomal synthesis dependent proteolipid showing a high affinity for phosphate

Cited by 44 publications

References 24 publications

Inhibition of Anion Transport in Corn Root Protoplasts

Inhibition of Anion Transport in Corn Root Protoplasts

Existence of two different species of mitochondrially translated proteolipids in ATPase complex of yeast mitochondria

Amino acid sequence of a new mitochondrially synthesized proteolipid of the ATP synthase of Saccharomyces cerevisiae.

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