Title

Interaction of CPa-1 with the Manganese-Stabilizing Protein of Photosystem II: Identification of Domains on CPa-1 Which Are Shielded from N-Hydroxysuccinimide Biotinylation by the Manganese-Stabilizing Protein

Document Type

Article

Publication Date

2-1-1992

Abstract

The structural organization of photosystem II proteins has been investigated by use of the amino group-labeling reagent 7V-hydroxysuccinimidobiotin (NHS-biotin) and calcium chloride-washed photosystem II membranes. We have previously shown that the presence of the extrinsic, manganese-stabilizing protein on photosystem II membranes prevents the modification of lysyl residues located on the chlorophyll protein CPa-1 (CP-47) by NHS-biotin [Bricker, T.M., Odom, W.R., & Queirolo, C.B. (1988) FEBS Lett. 231, 111-117]. Upon removal of the manganese-stabilizing protein by calcium chloride-washing, CPa-1 can be specifically modified by treatment with NHS-biotin. Preparative quantities of biotinylated CPa-1 were subjected to chemical cleavage with cyanogen bromide. Two major biotinylated peptides were identified with apparent molecular masses of 11.8 and 15.7 kDa. N-terminal sequence analysis of these peptides indicated that the 11.8-kDa peptide was 232G-330M and that the 15.7-kDa peptide was 360P-508V. The 15.7-kDa CNBr peptide was subjected to limited tryptic digestion. The two smallest tryptic fragments identified migrated at apparent molecular masses of 9.1 (nonbiotinylated) and 7.5 kDa (biotinylated). N-terminal sequence analysis and examination of the predicted amino acid sequences of these peptides suggest that the 9.1-kDa fragment was 422R-508V and that the 7.5-kDa fragment was 360P-421A. These results strongly suggest that two NHS-biotinylated domains, 304K-321K and 389K-419K, become exposed on CPa-1 when the manganese-stabilizing protein is removed by CaCl2treatment. Both of these domains lie in the large extrinsic loop E of CPa-1. © 1992, American Chemical Society. All rights reserved.

Publication Source (Journal or Book title)

Biochemistry

First Page

11059

Last Page

11064

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