Dissection of functional domains in phage fd adsorption protein

Stengele, Irina; Bross, Peter; Garces, Xaver; Giray, Jeanette; Rasched, Ihab

doi:10.1016/0022-2836(90)90311-9

Cited by 111 publications

(30 citation statements)

References 33 publications

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“…Consequently, the peeled particle has about 20% of the negative charges of the native particle and its molecular mass is reduced by approximately 30%, ncvertheless it conserves the original shape of the virion. Peeled phage fd is non-infectious, because it has lost the minor coat protein responsible for host recognition and penetration (Stengele et al, 1990). It still bears negative charges (due to Glu20) and is water soluble.…”

Section: Discussionmentioning

confidence: 99%

Subtilisin removes the surface layer of the phage fd coat

Schwind¹,

Krämer²,

Kremser³

et al. 1992

European Journal of Biochemistry

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The major coat protein of native filamentous phage fd is vulnerable to digestion by subtilisin, but not by any of a number of other proteolytic enzymes. Degradation by the non-specific protease subtilisin occurs at specific sites in the N-terminal portion of g8p. The N-terminal part of the protein is considered to be the outer layer of a two-layered coat. Thus, subtilisin treatment results in a monolayered phage particle. These particles possess the morphology and stability of native phage fd. Furthermore, subtilisin proteolysis proved to be an efficient instrument in detecting variations in the topology of the g8p of related filamentous phages.

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Section: Discussionmentioning

confidence: 99%

Subtilisin removes the surface layer of the phage fd coat

Schwind¹,

Krämer²,

Kremser³

et al. 1992

European Journal of Biochemistry

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“…Mature g3p consists of 406 amino acid residues separated in several distinct regions (2). The N-terminal part can be divided into two different domains, N1 and N2, mediating penetration and adsorption during infection, respectively (1,30). The C-terminal part is responsible for the interaction with g6p, thereby anchoring the g3p in the membrane of the phage coat (1,14,21).…”

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confidence: 99%

“…The first linker consists of repeats of the sequence Glu-Gly-Gly-Gly-Ser, and the second linker consists of repeats of the sequence Gly-Gly-Gly-Ser. There have been reports showing that the first linker can have an effect on the outer membrane, resulting in ␤-lactamase leakage, impaired F pili, and tolerance to certain colicins (4,30). Apart from these observations, no proper function has so far been assigned to these regions, which are believed to mainly convey flexibility to the other domains of g3p.…”

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The Phage Infection Process: a Functional Role for the Distal Linker Region of Bacteriophage Protein 3

Nilsson

Malmborg

Borrebaeck

2000

J Virol

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The filamentous bacteriophage infects Escherichia coli by interaction with the F pilus and the TolQRA complex. The virus-encoded protein initiating this process is the gene 3 protein (g3p). The g3p molecule can be divided into three different domains separated by two glycine-rich linker regions. Though there has been extensive evaluation of the importance of the diverse domains of g3p, no proper function has so far been assigned to these linker regions. Through the design of mutated variants of g3p that were displayed on the surface of bacteriophage, we were able to elucidate a possible role for the distal glycine-rich linker region. A phage that displayed a g3p comprised of only the N1 domain, the first linker region, and the C-terminal domain was able to infect cells at almost the same frequency as the wild-type phage. This infection was proven to be dependent on the motif between amino acid residues 68 and 86 (i.e., the first glycine-rich linker region of g3p) and on F-pilus expression.Filamentous bacteriophage is a nonlytic DNA virus that infects Escherichia coli cells harboring an F episome (19,20,26). In order for bacteriophage infection to occur, including the translocation of the single-stranded phage DNA over the cell membrane, the expression of two different receptor systems in the gram-negative host cell is critical. The primary receptor, i.e., the F pilus, mediates the adsorption of the phage to the bacterial cell (6,12). This interaction can take place at a long distance from the cell surface, since the pilus molecule extrudes away from the bacteria. Through the depolymerization of the pilus, the bacteriophage is brought close to the cell surface of the host, where the interaction with the coreceptor takes place (5, 31). The coreceptor, i.e., the TolQRA complex, is localized in the inner membrane and the periplasmic space and is attached to the inside of the outer membrane (9, 16). It was recently shown that it is the C-terminal part of the TolA domain that interacts with the bacteriophage adsorption protein (7,27). Upon binding to the TolA domain, the viral DNA translocates into the host cell by an as-yet-unknown mechanism.Expression of the gene 3 viral adsorption protein, g3p, on the surface of the bacteriophage is essential for normal infection to occur (1). The minor coat protein g3p is located together with g6p in three to five copies at one end of the filamentous phage (10). Mature g3p consists of 406 amino acid residues separated in several distinct regions (2). The N-terminal part can be divided into two different domains, N1 and N2, mediating penetration and adsorption during infection, respectively (1, 30). The C-terminal part is responsible for the interaction with g6p, thereby anchoring the g3p in the membrane of the phage coat (1, 14, 21). Furthermore, the g3p has a crucial role in the assembly of the filamentous phage, since in the absence of g3p, the proper termination of the assembly and the following release of the phage will not take place (24).Two glycine-rich linker regions divid...

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“…2) [7]. The first two domains (N1-N2) have been shown to be responsible for the docking of the phage to the F-pili of male E. coli and subsequent penetration of the bacterial membrane [8, 9,10], while the C-terminal domain (CT) is presumed to have a structural role, making contact with the DNA and forming the tip of the phage particle [11].…”

Section: Introductionmentioning

confidence: 99%

Co‐selection of cognate antibody‐antigen pairs by selectively‐infective phages

et al. 1995

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We have developed a chloramphenicol resistant derivative of fd phage with which cognate pairs of antibodies and antigens can be selected. The phage genome encodes a fusion of single-chain antibody to the C-terminal domain of glIIp, rendering the phage non-infective. The antigen fused to the N-terminal domains of glIIp is encoded in the same phage genome. Antigen and antibody fusion interact with each other in the periplasm of the phage-producing cell, restoring infectivity. This system has a very low background and will allow simultaneous randomisation of antibody and antigen.

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Dissection of functional domains in phage fd adsorption protein

Cited by 111 publications

References 33 publications

Subtilisin removes the surface layer of the phage fd coat

Subtilisin removes the surface layer of the phage fd coat

The Phage Infection Process: a Functional Role for the Distal Linker Region of Bacteriophage Protein 3

Co‐selection of cognate antibody‐antigen pairs by selectively‐infective phages

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