Download
slide1 n.
Skip this Video
Loading SlideShow in 5 Seconds..
Supplemental Fig. S1 PowerPoint Presentation
Download Presentation
Supplemental Fig. S1

Supplemental Fig. S1

75 Vues Download Presentation
Télécharger la présentation

Supplemental Fig. S1

- - - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript

  1. Supplemental Fig. S1 a) b) PsAPY1 (AB071369) 1:MEFLIKLIT-FLLFSMPAITSSQYLGNNLLTSRKIFLKQEEISSYAVVFDAGSTGSRIHV 59 MtAPY1;1 (AY180377) 1:MEFLITLITTVLLLLMPAITSSQYLGNNLLTNRKIFQKQETISSYAVVFDAGSTGSRIHV 60 ***** *** ** **************** **** *** ******************* PsAPY1 (AB071369) 60:YHFNQNLDLLHIGKGVEYYNKITPGLSSYANNPEQAAKSLIPLLEQAEDVVPDDLQPKTP 119 MtAPY1;1 (AY180377) 61:YHFDQNLDLLHIGKDVEFFNKITPGLSSYANDPEQAAKSLIPLLQQAENVVPIDLHHKTP 120 *** ********** ** ************ ************ *** *** ** *** PsAPY1 (AB071369) 120:VRLGATAGLRLLNGDASEKILQSVRDMLSNRSTFNVQPDAVSIIDGTQEGSYLWVTVNYA 179 MtAPY1;1 (AY180377) 121:IRLGATAGLRLLNGDASEKILQAVRDMFSNRSTFNVQPDAVSIIDGTQEGSYLWVTVNYA 180 ********************* **** ******************************** PsAPY1 (AB071369) 180:LGNLGKKYTKTVGVIDLGGGSVQMAYAVSKKTAKNAPKVADGDDPYIKKVVLKGIPYDLY 239 MtAPY1;1 (AY180377) 181:LGNLGKKYTKTVGVMDLGGGSVQMAYAVSKKTAKNAPKVADGVDPYIKKLVLKGKPYDLY 240 ************** *************************** ****** **** ***** PsAPY1 (AB071369) 240:VHSYLHFGREASRAEILKLTPRSPNPCLLAGFNGIYTYSGEEFKATAYTSGANFNKCKNT 299 MtAPY1;1 (AY180377) 241:VHSYLHFGREASRAEIMKVTRSSPNPCLLAGFDGTYTYAGEEFKAKAPASGANFNGCKKI 300 **************** * * ********** * *** ****** * ****** ** PsAPY1 (AB071369) 300:IRKALKLNYPCPYQNCTFGGIWNGGGGNGQKNLFASSSFFYLPEDTGMVDASTPNFILRP 359 MtAPY1;1 (AY180377) 301:IRKALKLNYPCPYQNCTFGGIWNGGGGNGQKHLFASSSFFYLPEDVGMVDPKTPNFKIRP 360 ******************************* ************* **** **** ** PsAPY1 (AB071369) 360:VDIETKAKEACALNFEDAKSTYPFLDKKNVASYVCMDLIYQYVLLVDGFGLDPLQKITSG 419 MtAPY1;1 (AY180377) 361:VDLVSEAKKACALNFEDAKSTYPFLAKKNIASYVCMDLIYQYVLLVDGF--DPLQEITSG 418 ** ** **************** *** ******************* **** **** PsAPY1 (AB071369) 420:KEIEYQDAIVEAAWPLGNAVEAISALPKFERLMYFV 455 MtAPY1;1 (AY180377) 419:KEIEYQDAVLEAAWPLGNAVEAISSLPKFERMMYFV 454 ******** ************** ****** **** Fig. 1 Apyrase protein can be detected with the antibody generated against a mixture of synthetic peptides conserved in PsAPY1 and MtAPY1;1. a) Alignment of PsAPY1 (AB071369) and MtAPY1;1 (AY180377) proteins. Synthesized peptides (underlined) were injected into a rabbit to produce an antigen-specific antibody. b) Cell wall proteins from pea epicotyl tissues were separated on 12% SDS-PAGE gel and subjected to a western blot with the antibody. The blot was visualized with a ChemiDoc XRS (Bio-Rad), using ECL Plus Western Blotting Detection Reagents (GE Healthcare Life Science). 50kDa - *

  2. Supplemental Fig. S2 a) 2138.1500R.LAFHYIGLDDPEKDHVLR.Y 1412.7600 R.YETHPTLVSIPR.K 990.4900 K.FHIGFDVR.A 1591.9000 R.AGIVISLASIYDLEK.H 2472.2500 K.GYISELFVPYQDPTEEFYFK.T 1861.1000 K.ITIGLAPAELVVVNPNIK.T 2272.3100 R.LIPAIPAHPLLTEDDYPQIR.G 1260.6200 K.WAGGLYVDHSR.G b) Fig. 2 Identification of p74 protein as a copper amine oxidase. a) MALDI-TOF-MS spectrum data for the tryptic peptides. b) Amino acid sequence of copper amine oxidase (CuAO) of pea. Matched peptides are underlined. 1 MASTTTMRLA LFSVLTLLSF HAVVSVTPLH VQHPLDPLTK EEFLAVQTIV 51 QNKYPISNNR LAFHYIGLDD PEKDHVLRYE THPTLVSIPR KIFVVAIINS 101 QTHEILINLR IRSIVSDNIH NGYGFPILSV DEQSLAIKLP LKYPPFIDSV 151 KKRGLNLSEI VCSSFTMGWF GEEKNVRTVR LDCFMKESTV NIYVRPITGI 201 TIVADLDLMK IVEYHDRDIE AVPTAENTEY QVSKQSPPFG PKQHSLTSHQ 251 PQGPGFQING HSVSWANWKF HIGFDVRAGI VISLASIYDL EKHKSRRVLY 301 KGYISELFVP YQDPTEEFYF KTFFDSGEFG FGLSTVSLIP NRDCPPHAQF 351 IDTYVHSANG TPILLKNAIC VFEQYGNIMW RHTENGIPNE SIEESRTEVN 401 LIVRTIVTVG NYDNVIDWEF KASGSIKPSI ALSGILEIKG TNIKHKDEIK 451 EDLHGKLVSA NSIGIYHDHF YIYYLDFDID GTHNSFEKTS LKTVRIKDGS 501 SKRKSYWTTE TQTAKTESDA KITIGLAPAE LVVVNPNIKT AVGNEVGYRL 551 IPAIPAHPLL TEDDYPQIRG AFTNYNVWVT AYNRTEKWAG GLYVDHSRGD 601 DTLAVWTKQN REIVNKDIVM WHVVGIHHVP AQEDFPIMPL LSTSFELRPT 651 NFFERNPVLK TLSPRDVAWP GCSN

  3. Supplemental Fig. S3 Fig. 3 Ecto-apyrase PsAPY1 from pea interacts with a copper amine oxidase PsCuAO in yeast. The coding regions for PsAPY1 (GenBank accession AB071369; Kawahara et al. 2003) and PsCuAO (GenBank/ENBL data bank accession L39931; Tipping andMcPherson 1996) were amplified by PCR and ligated into pGBKT7 as a bait and pGADT7 as a prey, respectively, and tested for the interaction using Saccharomyces cerevisiae strain AH109, according to the manufacturer’s protocol. The transformed S.cerevisiae expressing both PsAPY1 and PsCuAO successfully grows on selective media (SD/-His/-Lue/-Trp). Note that the interaction was detected by reciprocal bait–prey pairing. Bait - - PsAPY1 PsCuAO PsAPY1 PsCuAO - - Prey PsCuAO PsAPY1 PsAPY1 PsCuAO SD (-His/-Lue/-Trp) YPDA