Aggregation-related conformational change of the membrane-associated coat protein of bacteriophage M13

Spruijt, Ruud B.; Wolfs, C.J.A.M.; Hemminga, M.A.

doi:10.1021/bi00449a030

Cited by 89 publications

(134 citation statements)

References 44 publications

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“…complexes has previously been shown to be essentially the same as the CD spectrum of pVIII in lipid bilayers [23,26--28]. These CD spectra are consistent with about 50-60% helix [26][27][28], which agrees with a Raman estimate of 55% helix using the amide I band [6]. The Raman studies indicated that the SDSpVIII complexes contain a mixture of helix, turns and random, with very little sheet [6], suggesting a transmembrane helix and nonhelical ends for the lnembranebound form.…”

Section: And Roman Spectrasupporting

confidence: 68%

A model for fd phage penetration and assembly

et al. 1991

FEBS Letters

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Below 15°C, chloroform causes fd phage to contract to I‐forms, which are compact structures about 1/3 as long as the original phage. Above 15°C, chloroform causes I‐forms to contract to even more compact spheroidal S‐forms. Here we show that the coat protein structure in I‐forms is the same as the protein structure in the phage and the protein structure in S‐forms is the same as the protein structure in bilayers. The conversions from fda˚I‐forms→S‐forms are therefore suggested to mimic steps in fd penetration. The same conversions, in reverse order, are suggested to mimic steps in fd assembly.

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Section: And Roman Spectrasupporting

confidence: 68%

A model for fd phage penetration and assembly

et al. 1991

FEBS Letters

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“…M 13 coat protein when reconstituted in phospholipid systems is able to adopt different aggregation states depending on its lipidic environment and history of preparation (Spruijt et al, 1989;Spruijt & Hemminga, 1991). Therefore, the aggregation state of the protein was checked in the various reconstituted lipid-protein systems.…”

Section: Resultsmentioning

confidence: 99%

“…The coat protein in a predominant /3-sheet conformation appears to be aggregated in an irreversible way (/3-polymeric protein). The presence of either one of these M l3 coat protein forms depends on the protein purification procedure and reconstitu tion conditions (Spruijt et al, 1989;Spruijt & Hemminga, 1991). M l3 coat protein in the a-oligomeric form is assumed to be the biological active form in membraneassociated processes (Spruijt & Hemminga, 1991;Hemminga et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

FT-IR spectroscopy of the major coat protein of M13 and Pf1 in the phage and reconstituted into phospholipid systems

Wolkers¹,

Haris²,

Pistorius³

et al. 1995

Biochemistry

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FT-IR spectroscopy has been applied to study the secondary structure of the major coat protein of Pfl and M l 3 as present in the phage and reconstituted in DOPG and mixed DOPC/DOPG (4/1) bilayers. Infrared absorbance spectra of the samples were examined in dehydrated films and in suspensions of DiO and H2O. The secondary structure of the coat protein is investigated by second-derivative analysis, Fourier self-deconvolution, and curve fitting of the infrared bands in the amide I region (1600-1700 cm "1), is found that, in dehydrated films o f P fl and M13 phage, the amide I region contains three bands located at about 1633, 1657, and 1683 c m "1, that are assigned to hydrogen-bonded turn, a-helix/random coil, and non-hydrogen-bonded turn, respectively. From a comparison of the infrared spectra in dehydrated film with those in aqueous suspension, the percentages of secondary structure were found with an accuracy of about ±5%. For the coat protein of P fl phage, the FT-IR quantification gives 69% a-helix conformation, 19% turn structure, and 12% random coil structure. For P fl coat protein in the membrane-embedded state, the amount of a-helix is 57%, whereas 42% is in a turn structure and 1% in a random coil structure.

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“…15NH,Cl and ["C]glucose were used as the bole carbon and nitrogen sources to obtain uniformly "Nand "C-labelled M13 phages. The isolation and purification of the phages were performed as described earlier (Konings, 1980;Spruyt et al, 1989). Separation of gVIIIp from the phage particles was carried out by phenol extraction (Knippers and Hoffman-Berling, 1966;Konings et al, 1970).…”

Section: Methodsmentioning

confidence: 99%

NMR Studies of the Major Coat Protein of Bacteriophage M13

Papavoine

Aelen

Konings

et al. 1995

European Journal of Biochemistry

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The membrane-bound form of the major coat protein (gV11Ip) of bacteriophage M I 3 has been studied using nuclear magnetic resonance spectroscopy. As membrane mimetics, we used dodecylphosphocholine (DodPCho) detergent micelles to solubilize the protein. We were able to nearly completely assign all resonances of the protein solubilized in DodPCho micelles by using both homonuclear and heteronuclear multidimensional experiments. Based on the patterns of the nuclear Overhauser enhancements and the chemical shifts of the resonances, we deduced the secondary structure of the protein. Additional structural information was obtained from amide proton exchange data and J-coupling constants. The protein consists of two a-helices which are connected by a hinge region around residue 21. From spin-label experiments, the location of the protein relative to the DodPCho micelles was determined. One, hydrophobic, helix spans the micelle, and another, amphipathic, helix, is located beneath the surface of the micelle. For gVTTTp in SDS micelles, we found a micellar structure which is distorted near the C-terminus of the protein; whereas for DodPCho micelles, both distorted and regular elliptical micelles occur. This distortion is probably due to the interaction of the positively charged lysine side chains with the negatively charged head group of the detergent molecules.Keywords: major coat protein; bacteriophage M I 3 ; NMR; dodecylphosphocholine; structural analysis.Our knowledge of biological processes involving membranebound proteins is limited in comparison with those involving cytoplasmic proteins. This is due, in large part, to the lack of structural information caused by difficulties in applying the methods of X-ray diffraction and NMR to membrane-bound proteins. Membrane-associated proteins are more difficult to crystallize than globular proteins. Multidimensional high-resolution NMR methods, which work well in the elucidation of the structures of small water-soluble proteins (Wiithrich, 1986) are less well-suited for studying large, slowly reorienting complexes, like membrane-bound proteins. To obtain structural information for these proteins with these techniques, model systems, mostly in the form of detergent micelles, are used as membrane mimetics. In high-resolution NMR, glucagon (Braun et al., 1981) and melittin (Brown et al., 1982) were the initial model systems Correspondence to F. J. M. Van de Ven,

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Aggregation-related conformational change of the membrane-associated coat protein of bacteriophage M13

Cited by 89 publications

References 44 publications

A model for fd phage penetration and assembly

A model for fd phage penetration and assembly

FT-IR spectroscopy of the major coat protein of M13 and Pf1 in the phage and reconstituted into phospholipid systems

NMR Studies of the Major Coat Protein of Bacteriophage M13

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