Crystallization and preliminary X-ray crystallographic analysis of Mirabilis antiviral protein

Miyano, Masashi; Appelt, Krzystof; Arita, Masatoshi; Habuka, Noriyuki; Kataoka, Jiro; Ago, Hideo; Tsuge, Hideaki; Noma, Masana; Ashford, V.; Xuong, Nguyen‐Huu

doi:10.1016/0022-2836(92)90142-7

Cited by 8 publications

(4 citation statements)

References 13 publications

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“…The rod-like crystals of Y72F mutant, grown according to Miyano et al (1992), belong to space group P3121 (or P3221) with a = b = 104.1 and c--134.3 A. The crystals are isomorphous with the wildtype crystals, but diffract to higher resolution and exhibit much less twinning.…”

Section: Resultsmentioning

confidence: 99%

Improved crystals of the toxic protein MAP by protein engineering towards the host specificity

Ago¹,

Habuka²,

Kataoka³

et al. 1994

Acta Cryst D

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Mirabilis anti-viral protein (MAP) is a ribosomeinactivating protein from Mirabilis jalapa L. Since MAP is effective over a broad spectrum of species, the protein is difficult to express in heterologous hosts such as Escherichia coli. Recently, we obtained a MAP mutant, Y72F which exhibits a lower (1/100) activity against E. coli ribosomes while retaining almost full activity against mammalian cells . J. Biol. Chem. 267,[7758][7759][7760]. For the crystallographic studies, the Y72F MAP expression vector with an OmpA leading sequence was constructed and expressed in E. coli. The Y72F MAP mutant was then isolated and purified from the cell culture medium. Crystals were grown using the crystallization conditions for the native MAP crystals . J. Mol. Biol. 226,[281][282][283]: 50% ammonium sulfate containing 50 mM ammonium citrate and 2 mM adenine sulfate, pH 5.4. The crystals belong to space group P3~21 (or P3221) with a = b = 104.1 and c = 134.3 A. The crystals are isomorphous with the wild-type crystals but diffract to higher resolution. Imaging-plate photographs of the Y72F mutant showed sharp intense spots without the streaking observed in the native crystals.

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Section: Resultsmentioning

confidence: 99%

Improved crystals of the toxic protein MAP by protein engineering towards the host specificity

Ago¹,

Habuka²,

Kataoka³

et al. 1994

Acta Cryst D

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show abstract

“…The X-ray structure of several type 1 RIPs, as -and -momorcarin (Husain et al, 1994;Yuan et al, 1999), PAP (Mozingo et al, 1993), -trichosanthin (Gao et al, 1994), cucurmarin (Chen et al, 2000), MAP30 (Miyano et al, 1992), gelonin (Hosur et al, 1995), bryodin 1 (Gawlak et al, 1997), saporin-S6 (Savino et al, 2000), and -luYn (Ma et al, 2003) and type 2 RIPs, as ricin (Rutenber et al, 1991), shiga toxin (Fraser et al, 1994), abrin A (Tahirov et al, 1995), ebulin 1 (Pascal et al, 2001), and viscumin (Sweeney et al, 1993) is known (reviewed in Robertus and Monzingo, 2004). It appears that widely distributed RIPs of both types must originate from the same ancestor and assume the same "RIP fold.…”

Section: Introductionmentioning

confidence: 98%

The 1.4Å structure of dianthin 30 indicates a role of surface potential at the active site of type 1 ribosome inactivating proteins

Fermani

Falini

Ripamonti

et al. 2005

Journal of Structural Biology

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“…Crystallisation and preliminary X‐ray crystallographic analysis of Mirabilis antiviral protein has been reported 18…”

Section: Introductionmentioning

confidence: 99%

Reliable sequence determination of ribosome- inactivating proteins by combining electrospray mass spectrometry and Edman degradation

et al. 2001

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The primary structure of saporin-S9 and MAP-S, two type-1 ribosome-inactivating proteins isolated from the seeds of Saponaria officinalis L. and Mirabilis jalapa, respectively, was determined using a combined approach based on Edman degradation and electrospray ionization mass spectrometry (ESMS). Saporin-S9 has 253 amino acids with a calculated molecular mass of 28,492.99, which is in good agreement with that determined by ESMS (28 495 +/- 2 Da). Unlike other saporins with known primary structure, saporin-S9 contains four histidinyl residues (positions 111, 121, 216 and 248). By comparing the amino acid sequence of saporin-S9 with that of saporin-S6, we found 22 amino acid substitutions (8.7%), 13 of which are conservative and nine non-conservative. The residues known to be involved in the definition of the active site and with RNA base recognition are conserved. The four histidinyl residues and especially Lys for Gln203 contribute to the higher calculated pI value (10.17) of saporin-S9 compared with saporin-S6 (9.98). MAP-S contains 250 amino acid residues with a calculated molecular mass of 27,789.49, in good agreement with that determined by ESMS (27,789 +/- 2). Cys36 and Cys220 form a disulphide bridge and only four amino acid residues are different from the amino acid sequence of MAP, isolated from the roots of the same plant, i.e. Leu34 (Glu), Ile161 (Leu), Asp185 (Glu) and Asp191 (Glu) (in parentheses, the residues present in MAP). The reported approach can provide rapid and reliable sequence screening in the analysis of homologous proteins, including the presence of disulphide bridges.

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Crystallization and preliminary X-ray crystallographic analysis of Mirabilis antiviral protein

Cited by 8 publications

References 13 publications

Improved crystals of the toxic protein MAP by protein engineering towards the host specificity

Improved crystals of the toxic protein MAP by protein engineering towards the host specificity

The 1.4Å structure of dianthin 30 indicates a role of surface potential at the active site of type 1 ribosome inactivating proteins

Reliable sequence determination of ribosome- inactivating proteins by combining electrospray mass spectrometry and Edman degradation

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