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. 1992 Dec;11(12):4401–4409. doi: 10.1002/j.1460-2075.1992.tb05540.x

Identification of two structurally related proteins involved in proteolytic processing of precursors targeted to the chloroplast.

J E Oblong 1, G K Lamppa 1
PMCID: PMC557014  PMID: 1385116

Abstract

Two proteins of 145 and 143 kDa were identified in pea which co-purify with a chloroplast processing activity that cleaves the precursor for the major light-harvesting chlorophyll binding protein (preLHCP). Antiserum generated against the 145/143 kDa doublet recognizes only these two polypeptides in a chloroplast soluble extract. In immunodepletion experiments the antiserum removed the doublet, and there was a concomitant loss of cleavage of preLHCP as well as of precursors for the small subunit of Rubisco and the acyl carrier protein. The 145 and 143 kDa proteins co-eluted in parallel with the peak of processing activity during all fractionation procedures, but they were not detectable as a homo- or heterodimeric complex. The 145 and 143 kDa proteins were used separately to affinity purify immunoglobulins; each preparation recognized both polypeptides, indicating that they are antigenically related. Wheat chloroplasts contain a soluble species similar in size to the 145/143 kDa doublet.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abad M. S., Clark S. E., Lamppa G. K. Properties of a Chloroplast Enzyme that Cleaves the Chlorophyll a/b Binding Protein Precursor : Optimization of an Organelle-Free Reaction. Plant Physiol. 1989 May;90(1):117–124. doi: 10.1104/pp.90.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abad M. S., Oblong J. E., Lamppa G. K. Soluble Chloroplast Enzyme Cleaves preLHCP Made in Escherichia coli to a Mature Form Lacking a Basic N-Terminal Domain. Plant Physiol. 1991 Aug;96(4):1220–1227. doi: 10.1104/pp.96.4.1220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chitnis P. R., Harel E., Kohorn B. D., Tobin E. M., Thornber J. P. Assembly of the precursor and processed light-harvesting chlorophyll a/b protein of Lemna into the light-harvesting complex II of barley etiochloroplasts. J Cell Biol. 1986 Mar;102(3):982–988. doi: 10.1083/jcb.102.3.982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark S. E., Abad M. S., Lamppa G. K. Mutations at the transit peptide-mature protein junction separate two cleavage events during chloroplast import of the chlorophyll a/b-binding protein. J Biol Chem. 1989 Oct 15;264(29):17544–17550. [PubMed] [Google Scholar]
  5. Clark S. E., Lamppa G. K. Determinants for cleavage of the chlorophyll a/b binding protein precursor: a requirement for a basic residue that is not universal for chloroplast imported proteins. J Cell Biol. 1991 Aug;114(4):681–688. doi: 10.1083/jcb.114.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gavel Y., von Heijne G. A conserved cleavage-site motif in chloroplast transit peptides. FEBS Lett. 1990 Feb 26;261(2):455–458. doi: 10.1016/0014-5793(90)80614-o. [DOI] [PubMed] [Google Scholar]
  7. Geli V., Yang M. J., Suda K., Lustig A., Schatz G. The MAS-encoded processing protease of yeast mitochondria. Overproduction and characterization of its two nonidentical subunits. J Biol Chem. 1990 Nov 5;265(31):19216–19222. [PubMed] [Google Scholar]
  8. Hawlitschek G., Schneider H., Schmidt B., Tropschug M., Hartl F. U., Neupert W. Mitochondrial protein import: identification of processing peptidase and of PEP, a processing enhancing protein. Cell. 1988 Jun 3;53(5):795–806. doi: 10.1016/0092-8674(88)90096-7. [DOI] [PubMed] [Google Scholar]
  9. Lamppa G. K., Abad M. S. Processing of a wheat light-harvesting chlorophyll a/b protein precursor by a soluble enzyme from higher plant chloroplasts. J Cell Biol. 1987 Dec;105(6 Pt 1):2641–2648. doi: 10.1083/jcb.105.6.2641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lamppa G. K., Morelli G., Chua N. H. Structure and developmental regulation of a wheat gene encoding the major chlorophyll a/b-binding polypeptide. Mol Cell Biol. 1985 Jun;5(6):1370–1378. doi: 10.1128/mcb.5.6.1370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Laue T. M., Rhodes D. G. Determination of size, molecular weight, and presence of subunits. Methods Enzymol. 1990;182:566–587. doi: 10.1016/0076-6879(90)82045-4. [DOI] [PubMed] [Google Scholar]
  12. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  13. Oblong J. E., Lamppa G. K. Precursor for the light-harvesting chlorophyll a/b-binding protein synthesized in Escherichia coli blocks import of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem. 1992 Jul 15;267(20):14328–14334. [PubMed] [Google Scholar]
  14. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  15. Ou W. J., Ito A., Okazaki H., Omura T. Purification and characterization of a processing protease from rat liver mitochondria. EMBO J. 1989 Sep;8(9):2605–2612. doi: 10.1002/j.1460-2075.1989.tb08400.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pain D., Kanwar Y. S., Blobel G. Identification of a receptor for protein import into chloroplasts and its localization to envelope contact zones. Nature. 1988 Jan 21;331(6153):232–237. doi: 10.1038/331232a0. [DOI] [PubMed] [Google Scholar]
  17. Perry S. E., Buvinger W. E., Bennett J., Keegstra K. Synthetic analogues of a transit peptide inhibit binding or translocation of chloroplastic precursor proteins. J Biol Chem. 1991 Jun 25;266(18):11882–11889. [PubMed] [Google Scholar]
  18. Pollock R. A., Hartl F. U., Cheng M. Y., Ostermann J., Horwich A., Neupert W. The processing peptidase of yeast mitochondria: the two co-operating components MPP and PEP are structurally related. EMBO J. 1988 Nov;7(11):3493–3500. doi: 10.1002/j.1460-2075.1988.tb03225.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Robinson C., Ellis R. J. Transport of proteins into chloroplasts. Partial purification of a chloroplast protease involved in the processing of important precursor polypeptides. Eur J Biochem. 1984 Jul 16;142(2):337–342. doi: 10.1111/j.1432-1033.1984.tb08291.x. [DOI] [PubMed] [Google Scholar]
  20. Schnell D. J., Blobel G., Pain D. Signal peptide analogs derived from two chloroplast precursors interact with the signal recognition system of the chloroplast envelope. J Biol Chem. 1991 Feb 15;266(5):3335–3342. [PubMed] [Google Scholar]
  21. Stellwagen E. Gel filtration. Methods Enzymol. 1990;182:317–328. doi: 10.1016/0076-6879(90)82027-y. [DOI] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wasmann C. C., Reiss B., Bohnert H. J. Complete processing of a small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from pea requires the amino acid sequence Ile-Thr-Ser. J Biol Chem. 1988 Jan 15;263(2):617–619. [PubMed] [Google Scholar]
  24. Westwood J. T., Clos J., Wu C. Stress-induced oligomerization and chromosomal relocalization of heat-shock factor. Nature. 1991 Oct 31;353(6347):822–827. doi: 10.1038/353822a0. [DOI] [PubMed] [Google Scholar]
  25. Witte C., Jensen R. E., Yaffe M. P., Schatz G. MAS1, a gene essential for yeast mitochondrial assembly, encodes a subunit of the mitochondrial processing protease. EMBO J. 1988 May;7(5):1439–1447. doi: 10.1002/j.1460-2075.1988.tb02961.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yaffe M. P., Schatz G. Two nuclear mutations that block mitochondrial protein import in yeast. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4819–4823. doi: 10.1073/pnas.81.15.4819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yang M., Jensen R. E., Yaffe M. P., Oppliger W., Schatz G. Import of proteins into yeast mitochondria: the purified matrix processing protease contains two subunits which are encoded by the nuclear MAS1 and MAS2 genes. EMBO J. 1988 Dec 1;7(12):3857–3862. doi: 10.1002/j.1460-2075.1988.tb03271.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. von Heijne G., Steppuhn J., Herrmann R. G. Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem. 1989 Apr 1;180(3):535–545. doi: 10.1111/j.1432-1033.1989.tb14679.x. [DOI] [PubMed] [Google Scholar]

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