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Fig. 4. Import of proteins into Euglena chloroplasts requires intact chloroplasts and GTP hydrolysis but does not require an N-ethylmaleimide (NEM)-sensitive factor. An isolated 35S-labelled Golgi membrane fraction (M) was incubated in the light in the presence (+) or absence (–) of intact or osmotically lysed Euglena chloroplasts with (+) or without (–) addition of GTP{gamma}S and with (+) or without (–) pre-treatment of the chloroplast and cytoplasmic fractions with NEM. Fluorographs of [35S]pLHCPII immunoprecipitated from Percoll-purified chloroplasts show that conversion of pLHCPII to LHCPII and the 50 kDa partially processed pLHCPII polyprotein required intact chloroplasts. pLHCPII, LHCPII and the 50 kDa partially processed pLHCPII polyprotein were not recovered with chloroplasts in reactions containing GTP{gamma}S. LHCPII and the 50 kDa partially processed pLHCPII polyprotein were recovered with NEM treated reactions indicating Golgi to chloroplast transport and import required GTP hydrolysis but did not require an NEM-sensitive factor.





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