SupplementaryTable 1: Main Characteristics of patients and controls.
EOPD Patients(N=235) / LOPD Patients
(N=306) / Controls ≤50 (N=140) / Controls >50 (N=278)
% Male / 64% / 51% / 48% / 45%
%Family history / 34% / 31% / - / -
Mean Age / 41±8 / 66±9 / 45±8 / 72±10
Age range / 25-50 / 51-85 / 30-50 / 51-90
Supplementary Table 2: Primers used to amplified PARK2 coding region and 3’UTR
EXON / DIRECTION / 5’3’ / TEMPERATUREExon 1 / Forward / TCCGGGAGGATTACCCA / 59ºC
Reverse / ATCATGGTCACTGGGTAGGTG
Exon2 / Forward / ATGTTGCTATCACCATTTAAGGG / 62ºC
Reverse / AGATTGGCAGCGCAGGCGGCATG
Exon 3 / Forward / AATTGTGACCTGGATCAGCAG / 58ºC
Reverse / TGGACTTCCAGCTGGTGGT
Exon 4 / Forward / AGGCAATGTGTTAGTACACA / 58ºC
Reverse / ACAAGCTTTTAAAGAGTTCTTGT
Exon5 / Forward / ACATGTCTTAAGGAGTACATTT / 58ºC
Reverse / TCTCTAATTTCCTGGCAAACAGTG
Exon 6 / Forward / AGAGATTGTTTACTGTGGAAACA / 58ºC
Reverse / GAGTGATGCTATTTTTAGATCCT
Exon 7 / Forward / TGCCTTTCCACACTGACAGGTACT / 58ºC
Reverse / TCTGTTCTTCATTAGCATTAGAGA
Exon 8 / Forward / TGATAGTCATAACTGTGTGTAAG / 58ºC
Reverse / ACTGTCTCATTAGCGTCTATCTT
Exon 9 / Forward / GGGTGAAATTTGCAGTCAGT / 59ºC
Reverse / AATATAATCCCAGCCCATGTGCA
Exon 10 / Forward / ATTGCCAAATGCAACCTAATGTC / 58ºC
Reverse / TTGGAGGAATGAGTAGGGCATT
Exon 11 / Forward / ACAGGGAACATAAACTCTGATCC / 62ºC
Reverse / CAACACACCAGGCACCTTCAGA
Exon 12 / Forward / GTTTGGGAATGCGTGTTTT / 58ºC
Reverse / AGAATTAGAAAATGAAGGTAGACA
3’UTR- F1 / Forward / CACTGGTTCGACGTGTAGC / 59ºC
Reverse / TTGTGTCATCCGGAGGC
3’UTR- F2 / Forward / GTTTGTCCACACAAGACAGC / 60ºC
Reverse / TAGTTTGGAGAACCCACTGC
3’UTR- F3 / Forward / CCATTGACAAGGTGTTTCTC / 59ºC
Reverse / AATCATGAAAGGGAATGAAAG
3’UTR- F4 / Forward / GAATTACAGGGAAGAAATCAAG / 59ºC
Reverse / GGCTCCTCTTTAAAGATTCTG
3’UTR- F5 / Forward / GTGCCTATAAAGGAAACACGT / 59ºC
Reverse / TCCAATCCATGTCAACATAAGG
3’UTR- F6 / Forward / TCCCCAGGAGAGACCTTC / 60ºC
Reverse / TGAGACTCATGCCCTCAGT
3’UTR- F7 / Forward / TGTATTGTCTGACTTCTCAGG / 60ºC
Reverse / CCTCTGAGACGACGGGAC
3’UTR- F8 / Forward / TCCTAACTGCCTAAGATAACCC / 60ºC
Reverse / TCATGCCTGCCACCTCTC
3’UTR- F9 / Forward / GGTGGACAGATCAAAGTCTC / 60ºC
Reverse / CATGCAGATTGGGAAGGCG
3’UTR- F10 / Forward / CCATCATCCATAGCTTTCTTC / 59ºC
Reverse / CTTGAACTCCTGACCTCGTG
Supplementary Table 3. Summary of the EOPD patients with at least one PRNK mutation.
Index case / Mutation 1 / Mutation 2 / Age of onset / Family historyPJ94 / 255delA / - / 36 / Yes
PJ 182 / 255delA / - / 35 / No
Pj198 / 255delA / 255delA / 34 / No
PE231 / 255delA / 255delA / 36 / No
PJ234 / 255delA / 255delA / 23 / No
PJ235 / 255delA / 255delA / 33 / No
G06827 / 255delA / 255delA / 41 / Yes
G05186 / 255delA / 255delA / 26 / Yes
PJ271 / Met192Leu / - / 33 / No
PE488 / Met192Leu / - / 49 / No
PJ195 / Cys212Tyr / Cys212Tyr / 40 / No
G05043 / Cys212Tyr / Cys212Tyr / 22 / Yes
G06322 / Cys212Tyr / p.Gln34fs / 18 / No
G05230 / Thr240Met1 / - / 47 / Yes
PJ5 / Arg392_X394 / Arg392_X394 / 24 / Yes
PJ56 / Ala401_X567 / - / 48 / Yes
PJ209 / Ala401_X567 / - / 35 / No
PJ146 / Arg402Cys / - / 49 / No
PJ177 / Gly430Ser / - / 38 / No
PE660 / Pro437Leu / - / 49 / No
PJ74 / Pro437Leu / - / 40 / Yes
PJ148 / Val456Ile / - / 43 / No
G06500 / Trp445Stop / Trp445Stop / 40 / Yes
PJ103 / 255delA / Pro202Leu / 47 / Yes
PJ246 / C154_X176 / Ivs.11 -3C/G / 30 / Yes
G06472 / Deletion exon 4 / Thr240Met / 24 / Yes
G06598 / 255delA / Trp445_Stop / 17 / No
G06697 / Arg275Trp / 255delA / 29 / Yes
PJ122 / Deletion exon 3-5 / - / 39 / No
PJ91 / Duplication exon2 / - / 43 / Yes
PJ34 / Duplicationexon 3 / - / 26 / No
PJ157 / Duplication exon 3 / - / 36 / Yes
PJ171 / Duplication exon 3 / - / 29 / Yes
G06401 / Duplicationexon 3 / - / 18 / Yes
PJ92 / Duplication exon 4 / - / 40 / No
PJ145 / Duplication exon 4 / - / 49 / Yes
G05027 / Duplication exon 4 / - / 45 / Yes
G05049 / Duplication exon 4 / - / 45 / No
G06368 / Duplication exon 4 / - / 47 / Yes
PJ156 / Duplication exons 2-3 / - / 29 / Yes
PJ160 / Duplication exons 2-3 / - / 39 / No
PJ179 / Duplication exons 2-3 / - / 47 / No
PJ191 / Duplication exons 2-3 / - / 40 / No
PJ194 / Duplication exons 2-3 / - / 44 / No
Suplementary Table 4. PRKN 3’UTR variants, minor allele frequencies (MAF) and number of carriers of the MAF allele (value in parenthesis), in the EOPD and controls. The p value for the comparison of allele frequencies by chi square is also indicated.
Position / Variant id / MAF patients (n) / MAF controls (n) / p-valueControls / CEU*
*94A/G / Rs62637702 / 0.004 (2) / 0.026 (22) / 0.024 / 0.001
*93-94 Ins AC / new / <0.01 (1) / 0 (0) / - / 0.36
*94-95 delAC InsGC GC / - / <0.01 (4) / <0.01 (5) / - / 0.7
*180-238duplication / new / <0.01 (1) / 0 (0) / - / 0.36
*320 C/A / Rs71653628 / 0.04 (18) / 0.05 (43) / 0.03 / 0.34
*652 C/T / Rs71653629 / 0.14 (68) / 0.13 (116) / 0.15 / 0.5
*820 A/T / Rs74701717 / <0.01 (4) / <0.01 (8) / 0.003 / 0.7
*886-889 AGAT / Rs113233227 / <0.01 (4) / <0.01 (6) / - / 0.7
*1252 C/T / Rs77926621 / <0.01 (4) / <0.01 (8) / 0.003 / 1
*1296 A/G / Rs3734464 / 0.06 (31) / 0.05 (44) / 0.08 / 0.4
*1386 A/G / Rs140107485 / <0.01 (5) / <0.01 (8) / 0.005 / 1
*1694 C/T / Rs16892481 / <0.01 (3) / <0.01 (6) / 0.02 / 0.7
*1695 G/A / Rs77283740 / <0.01 (2) / <0.01 (6) / 0.003 / 0.7
*1818 G/T / new / 0.01 (1) / <0.01 (1) / - / 1
*2021 G/A / Rs1122470 / 0.08 (39) / 0.07 (59) / 0.06 / 0.5
*MAF in the EUR-Caucasian 1000genomes database (
Supplementary Figure 1: (A) SSCA patterns for a heterozygous *94A/G patient and three normal controls. (B) Sequence of a heterozygous patient for *94A/G polymorphism.
Supplementary Figure 2:MLPA study. Upper image corresponds to a deletion of exons3-5. Botton image correspond to an individual without any exon rearrangement.
Supplementary figure 3. Linkage disequilibrium (D´ values; haploview) between the
PRKN 3´UTR variants.