The Rubisco Large Subunit Binding Protein

• Published in: Philosophical transactions of the Royal Society of London. Series B, Biological sciences Biological sciences, 1986-10, Vol.313 (1162), p.419-428
• Main Authors: Musgrove, J. E., Ellis, R. J.
• Format: Article
• Language: English
• Subjects: Antibodies; Carrier proteins; Chloroplasts; Gels; Laboratory staining techniques; Oligomers; Plants; Protein precursors; Protein synthesis; Ribosomes
• ISSN: 0962-8436; 0080-4622; 1471-2970; 2054-0280
• DOI: 10.1098/rstb.1986.0048

Newly synthesized Rubisco large subunits made by isolated intact chloroplasts from Pisum sativum are bound non-covalently to another protein, termed the Rubisco large subunit binding protein. This protein is implicated in the assembly of Rubisco in higher plant chloroplasts. The binding protein has been purified from Pisum sativum in the form of an oligomer of relative molecular mass (M$_r$) about 720000. Analysis on polyacrylamide gels containing sodium dodecyl sulphate reveals equal amounts of two different types of subunit, termed alpha (M$_r$ about 61000) and beta (M$_r$ about 60000); thus the oligomer has the composition $\alpha_6\beta_6$. The alpha and beta subunits have been separated; their amino-terminal sequences are different, and antibodies raised against one subunit do not cross-react with the other subunit. Antibodies raised against the binding protein do not cross-react with the Rubisco large subunit, but do cross-react with polypeptides of M$_r$ about 60000 in extracts of chloroplasts from wheat, barley and tobacco, and in extracts of leucoplasts from castor-bean endosperm. The binding protein is made as a higher-molecular-mass precursor whan leaf polysomes are translated in a wheatgerm extract containing chloramphenicol, but is not synthesized by isolated intact chloroplasts. Thus the binding protein subunits are synthesized by cytoplasmic ribosomes and hence are likely to be encoded by nuclear genes. Etiolated Pisum plants contain binding protein, but exposure to light does not cause the same dramatic increase in amount that is seen in the case of Rubisco. Treatment of stromal extracts with Mg-ATP in the range 0.1-5.0 mM causes dissociation of the binding protein oligomer into monomeric subunits; CTP, GTP, UTP, AMP and cyclic AMP do not have this effect. Mg$^{2+}$ is required for dissociation but can be replaced by Ca$^{2+}$. Newly synthesized large subunits are released when the binding protein oligomer is dissociated, but re-attach when the dissociation is reversed by removal of ATP.