BioMed Cen al
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BMC Medical Gene ics
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Resea ch a icle
Se 80Ile mu a ion and a concu en P o25Leu a ian o he VHL
gene in an ex ended Hunga ian on Hippel-Lindau amily
A ila Pa ocs1, Pe e Ge gics2, Ka alin Balogh2, Miklos To h2, Fe enc Fazakas3,
Is an Liko4 and Ka oly Racz*2
Add ess: 1Molecula Medicine Resea ch G oup, Hunga ian Academy o Sciences and Semmelweis Uni e si y, Szen ki ályi 46, H-1088 Budapes ,
Hunga y, 22nd Depa men o Medicine, Facul y o Medicine, Semmelweis Uni e si y, Szen ki ályi 46, H-1088 Budapes , Hunga y, 3Medical and
Heal h Science Cen e , Uni e si y o Deb ecen, H-4200, Deb ecen, Hunga y and 4Rich e Gedeon LTD, Budapes , Hunga y
Email: A ila Pa ocs - [email p o ec ed]; Pe e Ge gics - ge pe @yahoo.com; Ka alin Balogh - bka us@ eemail.hu;
Miklos To h - o [email protected] e.hu; Fe enc Fazakas - wol i[email p o ec ed]e.hu; Is an Liko - I.Liko@ ich e .hu; Ka oly Racz* - [email p o ec ed].hu
* Co esponding au ho
Abs ac
Von Hippel-Lindau disease (VHL) is a a e au osomal dominan disease cha ac e ized by
de elopmen o cys ic and umo ous lesions a mul iple si es, including he b ain, spinal co d,
kidneys, ad enals, panc eas, epididymis and eyes. The clinical pheno ype esul s om molecula
abno mali ies o he VHL umo supp esso gene, mapped o human ch omosome 3p25-26. The
VHL gene encodes wo unc ionally ac i e VHL p o eins due o he p esence o wo ansla ional
ini ia ion si es sepa a ed by 53 codons. The majo i y o disease-causing mu a ions ha e been
de ec ed downs eam o he second ansla ional ini ia ion si e, bu he e a e con lic ing da a as o
whe he ew mu a ions loca ed in he i s 53 codons, such as he P o25Leu could ha e a
pa hogenic ole. In his pape we epo a la ge Hunga ian VHL ype 2 amily consis ing o 32
membe s in whom a disease-causing AGT80AAT (Se 80Ile) c.239G>A, p.Se 80Ile mu a ion, bu
no he concu en CCT25CTT (P o25Leu) c.74C>T, p.P o25Leu a ian co-seg ega ed wi h he
disease. To ou knowledge, he Se 80Ile mu a ion has no been p e iously desc ibed in VHL ype
2 pa ien s wi h high isk o pheoch omocy oma and enal cell cance . The e o e, his inding
ep esen s a no el geno ype-pheno ype associa ion and VHL kind eds wi h Se 80Ile mu a ion will
equi e ca e ul su eillance o pheoch omocy oma. We concluded ha he P o25Leu a ian is a
a e, neu al a ian , bu he p esence such a a e gene a ian may make gene ic counseling
di icul .
Backg ound
Von Hippel-Lindau disease (VHL) (OMIM n . 193300) is
a a e au osomal dominan mul i-o gan disease caused by
molecula abno mali ies o he VHL umo supp esso
gene [1]. Pa ien s wi h VHL a e a isk o de elopmen o
e inal, ce ebella , spinal, panc ea ic and enal heman-
gioblas omas, pulmona y and li e hemangiomas, clea -
cell enal ca cinomas, pheoch omocy omas, endolym-
pha ic sac umo s, mul iple enal, epididymal and panc e-
a ic cys s, cys adenomas o he epididymis and o he
b oad ligamen , and panc ea ic isle cell umo s [1-3].
Based on he p esence o absence o pheoch omocy oma,
wo main sub ypes o he VHL disease ha e been iden i-
ied. Pa ien s wi h VHL ype 1 a e a isk o de elop enal
Published: 16 Ap il 2008
BMC Medical Gene ics 2008, 9:29 doi:10.1186/1471-2350-9-29
Recei ed: 26 Oc obe 2007
Accep ed: 16 Ap il 2008
This a icle is a ailable om: h p://www.biomedcen al.com/1471-2350/9/29
© 2008 Pa ocs e al; licensee BioMed Cen al L d.
This is an Open Access a icle dis ibu ed unde he e ms o he C ea i e Commons A ibu ion License (h p://c ea i ecommons.o g/licenses/by/2.0),
which pe mi s un es ic ed use, dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed.
BMC Medical Gene ics 2008, 9:29 h p://www.biomedcen al.com/1471-2350/9/29
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cell ca cinoma and hemangioblas oma bu p edomi-
nan ly wi hou pheoch omocy oma, while hose wi h
VHL ype 2 may ha e all mani es a ions o he disease p e-
dominan ly including pheoch omocy oma. VHL ype 2
has been subdi ided in o sub ype 2A and 2B, wi h a low
and high isk o enal cell ca cinoma, espec i ely,
whe eas sub ype 2C is a pheoch omocy oma-only pheno-
ype. The p e alence o he VHL disease a ies be ween
1:39,000 in Ge many and 1:53,000 in Eas Anglia [4,5].
Mani es a ions o he disease show a iable exp ession
and se e al pa ien s may ha e only one mani es a ion [4].
The human VHL gene maps o ch omosome 3p25-26 [1].
As p edic ed by Knudson's wo-hi model, umo de elop-
men equi es inac i a ion o bo h copies; he i s hi , he
ge mline mu a ion o dele ion is ollowed by soma ic
al e a ions usually de ec ed as loss o he e ozygosi y. The
VHL gene has wo ansla ional ini ia ion si es sepa a ed
by 53 codons. O he wo p o eins encoded by he VHL
gene, he la ge one con ains 213 amino acids (pVHL30),
whe eas he sho e p o ein consis s o 160 amino acids
(pVHL18). Bo h p o eins a e unc ionally ac i e and hey
sha e he same mechanism o umo supp esso ac i i y
[6-8]. The VHL p o eins o m a mul ime ic complex wi h
Elongin B, Elongin C, Cul2 and Rbx1. This complex is
in ol ed in ubiqui in-dependen p o eolysis o la ge cel-
lula p o eins ia con olled deg ada ion o α-subuni s o
he he e odime ic ansc ip ion ac o hypoxia inducible
ac o (HIF) [9,10]. In addi ion, complexes con aining
pVHL, Elongin B, Elongin C, Cul2 and Rbx1 a ge p o-
eins a e in ol ed in cell-cycle egula ion o deg ada ion,
sugges ing ha pVHL has a possible ole in cell-cycle exi
[11]. In 1999, S ebbins and co-wo ke s p esen ed he
h ee-dimensional p o ein s uc u e o he VHL-ElonginC-
ElonginB complex. They iden i ied wo unc ionally ac i e
si es; he ElonginC binding si e in he α-helical domain
in ol ing esidues be ween amino acids 157 and 170, and
he HIF binding si e in he β-shee domain om esidue
91 o 113 [8].
The majo i y o VHL pa ien s ha e dele ions, inse ions, o
mu a ions downs eam o he second ansla ional ini ia-
ion si e loca ed a codon 54 o he VHL gene. In e es -
ingly, missense mu a ions encoding esidues o he
p o ein binding si es ha e been associa ed wi h VHL ype
2, whe eas VHL ype 1 is caused ei he by missense o non-
sense mu a ions a ec ing he hyd ophobic co e, o by pa -
ial gene dele ions which esul in a comple e de ec o
p o ein unc ion. I has been also demons a ed ha some
mu an pVHL, which a e associa ed wi h he ype 2C phe-
no ype, may impai ib onec in ma ix assembly while
hey e ain he abili y o down egula e HIF [12].
Because mu a ions a ec ing he i s 53 codons o he VHL
gene ha e no e ec on he s uc u e o he sho e VHL
p o ein, i seemed pa icula ly in e es ing o cla i y
whe he hese mu a ions could exe a pa hogenic e ec .
In one s udy he P o25Leu a ian has been associa ed
wi h a spo adic pheoch omocy oma [13]. bu o he s ud-
ies in ol ing a limi ed numbe o cases ailed o con i m
he pa hogenic ole o his a ian [14,15]. In his pape
we epo a la ge Hunga ian VHL ype 2 amily wi h a dis-
ease-causing AGT80AAT (Se 80Ile) c.239G>A, p.Se 80Ile
mu a ion and a concu en CCT25CTT (P o25Leu)
c.74C>T, p.P o25Leu a ian (iden i ica ion numbe :
s35460768 (dbSNP127) o he VHL gene. We show ha
in amily membe s he Se 80Ile mu a ion, bu no he
P o25Leu a ian co-seg ega ed wi h he disease. In o de
o con i m he pa hogenici y o he Se 80Ile mu a ion and
o es whe he he P o25Leu a ian migh be neu al, an
e olu iona y mul iple sequence alignmen analysis (MSA)
combined wi h a Align GVGD was.pe o med. Th ee-
dimensional modeling o he Ile80-mu an p o ein is also
p esen ed.
Pa ien s and me hods
Pa ien s and da a collec ion
A la ge Hunga ian VHL ype 2 amily spanning i e gene -
a ions and in ol ing 32 membe s (Fig 1) was e alua ed a
he 2nd Depa men o Medicine, Facul y o Medicine,
Semmelweis Uni e si y in Budapes , Hunga y. Ini ial
sc eening included medical his o y, physical examina ion,
abdominal ul asonog aphy, abdominal and b ain com-
pu ed omog aphy (CT) o magne ic esonance imaging
(MRI), oph halmologic examina ion, ou ine biochemi-
cal es ing and 24-h u ina y ca echolamine me aboli e
de e mina ions.
Sc eening o VHL gene mu a ions
W i en in o med consen was ob ained om all amily
membe s who pa icipa ed in he s udy. Genomic DNA
was ex ac ed om pe iphe al blood leukocy es using
DNA isola ion ki o mammalian blood samples (Boe-
h inge Mannheim Co po a ion, Indianapolis, IN). Mu a-
ion analysis o he VHL gene was pe o med as p e iously
desc ibed [16].
E olu iona y alignmen analysis o he VHL p o ein
Mul iple sequence alignmen (MSA) o p o ein sequences
was pe o med using 3D-co ee, a eely a ailable web-
based ool o aligning mul iple sequences [17]. A o al o
eigh sequences: Mus musculus (ENSMUSG0000003393
3), Ra us no egicus (ENSRNOG00000010258), Canis
amilia is (ENSCAFG00000005149), Homo sapiens (EN
ST00000256474), Gallus gallus (ENSGALG0000000136
78), and e olu iona y mo e dis an species including D o-
sophila melanogas e (CG13221_CG13221-RA), Xeno-
pus opicalis (ENSXET00000001448) and Taki ugu
ub ipes (SINFRUG00000121189) we e analysed.
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One popula me hod o measu ing biochemical dis-
ances be ween pai s o amino acids is he G an ham Di -
e ence (18), which akes in o accoun he composi ion,
pola i y and olume o mu an and wild- ype amino
acids. In his s udy, we ha e cons uc ed a VHL p o ein
MSA and combined a conse a ion sco e (GV) wi h a
measu e o biochemical di e ence be ween wild- ype and
mu an esidues wi h espec o he alignmen (GD). This
ex ension o he G an ham Di e ence, called Align
GVGD, has p e iously been success ully applied o BRCA1
and TP53 genes (19, 20). Align GVGD is eely a ailable
online (21).
Th ee-dimensional s uc u e modeling o he VHL p o ein
The coo dina es o he pVHL we e ob ained om
1 cb_a.pdb s uc u e ile [8]. The h ee-dimensional
image was gene a ed wi h Swiss-PdbViewe in combina-
ion wi h POV-Ray [22].
Resul s
Geno ype-pheno ype associa ions
The index pa ien was ope a ed o pheoch omocy oma
and enal cell ca cinoma diagnosed a he age o 34 y and
40 y , espec i ely. She also de eloped a b ain heman-
gioblas oma a he age o 42 y . Gene ic sc eening indi-
ca ed ha she had a he e ozygous AGT80AAT (Se 80Ile)
c.239G>A, p.Se 80Ile mu a ion o he VHL gene. The 70-
y -old mo he o he index pa ien p o ed o be no only
a gene ca ie o he Se 80Ile mu a ion, bu she also had
he CCT25CTT (P o25Leu) c.74C>T, p.P o25Leu a ian
(iden i ica ion numbe : s35460768 (dbSNP127)), bo h
in he e ozygous o ms. Howe e , he mo he showed no
clinical, biochemical o adiological e idence o VHL-
associa ed umo s. The absence o he P o25Leu a ian in
he index pa ien indica ed, ha he Se 80Ile mu a ion
and he P o25Leu a ian in he mo he we e p esen in
sepa a e alleles.
In addi ion o he index pa ien and he mo he , he
Se 80Ile mu a ion was de ec ed in o he 5 membe s o he
Pedig ee o a Hunga ian kind ed wi h on Hippel-Lindau diseaseFigu e 1
Pedig ee o a Hunga ian kind ed wi h on Hippel-Lindau disease.
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amily (III/7, III/8, IV/13, IV/14 and IV/16) (Fig 1), o
which all bu one membe (III/7, III/8, IV/13 and IV/14)
had clinical mani es a ions o he disease.
Impo an ly, pheoch omocy omas we e iden i ied in am-
ily membe s III/8 and IV/14, o whom, in pa ien IV/14
his was he i s sign o he disease mani es ed a he age
o 10 yea s and in ol ed bo h ad enals. O he ypical
mani es a ions included b ain hemangioblas omas in
amily membe s: III/7, III/8 and IV/13, e ina hemangiob-
las omas in amily membe s III/7, III/8, IV/13 and IV/14,
and enal cys s in amily membe IV/13. In addi ion, am-
ily his o y e ealed ha one sis e o he mo he o he
index pa ien (II/5) died due o bila e al enal cell ca cino-
mas. Ano he sis e o he mo he o he index pa ien
(II.4) died because o a me as a ic panc ea ic umo wi h-
ou his ological con i ma ion and, he e o e, his amily
membe was no conside ed as ha ing VHL. The age, clin-
ical mani es a ions, and he age a diagnosis o VHL-asso-
cia ed lesions in ca ie s o he Se 80Ile mu a ion and in
hose wi h clinically p o en VHL-associa ed umo s a e
summa ized in Table 1.
In addi ion o he mo he o he index pa ien , he
P o25Leu a ian was de ec ed on ano he amily membe
(III/6) (Fig 1), who was clinically heal hy.
The p e alence o he P o25Leu a ian was es ed in 16
membe s o o he 8 VHL amilies and in 39 pa ien s wi h
spo adic pheoch omocy omas, bu none o he pa ien s
had his a ian .
E olu iona y alignmen analysis o he VHL gene
As shown in Fig. 2., he Se a amino acid posi ion 80 ep-
esen s a highly conse ed amino acid esidue among di -
e en species, including a , mouse, dog, chicken and an
e olu iona y mo e dis an species, Xenopus opicalis. By
con as , he P o a amino acid posi ion 25 ailed o show
a conse ed pa e n. Using Align-GVGD c i e ia bo h a -
ian s we e p edic ed as neu al ul illing he c i e ia 0<GV
≤ 61,3 and GD>0 ( he calcula ed GV and GD sco es o
P o25Leu polymo phism we e 218.82 and 4.86 and o
Se 80Ile 219.56 and 3.24, espec i ely).
E ec o change o Se 80 o Ile80 on p o ein s uc u e
The h ee-dimensional model o he pVHL is a ailable
om esidue Me 54, which allowed us o analyse unc-
ional consequences o he Se 80Ile mu a ion using a
h ee-dimensional compu a ional modeling. The CPK
diag am in Fig. 3 illus a es bo h he wild- ype Se 80 and
he mu an Ile80 p o eins. The Se 80 esidue makes igh
junc ions wi h P o103 and Ile151 by hyd ogen-bonds
be ween he amino g oup o Se 80 and he ca boxyl oxy-
gen o P o103 and Ile151. The Se 80 esidue is loca ed on
he same side whe e ElonginC binds, and esidues P o81
and A g82 a e in ol ed in di ec binding. The Asn78 esi-
due om he β-domain, oge he wi h hyd ophobic co e
esidues (P o86, Phe76, Phe119, T p117 and Val130)
ha e impo an oles o s uc u al in eg i y o he β-sand-
wich [8]. A change o Se o Ile a posi ion 80 dis u bs his
in eg i y o he β-sandwich due o a change in he o ien a-
ion o he A g82 esidue, which dis u bs i s linke ole
and, consequen ly, he hyd ogen bond be ween A g82
and Leu153 disappea s. I has been al eady demons a ed
ha he A g82 esidue plays a cen al ole in s uc u al
in eg i y o he pVHL, as i makes signi ican hyd ogen-
bond con ac s wi h Leu153, Val155, Lys159 and A g161
and, h ough hese con ac s; i is in ol ed in in e ac ions
be ween pVHL and ElonginC [8].
Discussion
In his pape we desc ibe a la ge Hunga ian VHL ype 2
amily consis ing o 32 membe s, in whom a disease-caus-
ing AGT80AAT (Se 80Ile) c.239G>A, p.Se 80Ile mu a ion
Table 1: Clinical mani es a ions o VHL disease in ca ie s o he Se 80Ile mu a ion and/o he P o25Leu a ian and in hose wi h
clinically p o en VHL-associa ed umo s
Family membe
(age, yea s)
Renal cell
ca cinoma (age
a p esen a ion,
yea s)
B ain heman-
gioblas oma
(age a
p esen a ion,
yea s)
Pheoch omo-
cy oma (age a
p esen a ion,
yea s)
Re ina heman-
gioblas oma
(age a
p esen a ion,
yea s)
O he clinical
mani es a ions
Resul o
gene ic
sc eening
II/1 (70)-----P o25Leu Se 80Ile
III/6 (44)-----P o25Leu
II/5 (63) + bila e al (45) - - - ND
III/1* (42) + bila e al (40) + (42) + (34) - - Se 80Ile
III/7 (47) - + (37) - + (18) - Se 80Ile
III/8 (39) - + (38) + (39) + (29) - Se 80Ile
IV/13 (27) - + (22) - + (23) enal cys s (20) Se 80Ile
IV/14 (26) - - + bila e al (10) + (26) - Se 80Ile
IV/16 (16)-----Se 80Ile
*, index pa ien ; -, p esen ; +, absen ; ND, no de e mined (DNA o gene ic es ing was no a ailable)
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and a concu en CCT25CTT (P o25Leu) c.74C>T,
p.P o25Leu a ian we e iden i ied. Sc eening o amily
membe s indica ed ha 7 amily membe s had he
Se 80Ile mu a ion and 2 membe s had he P o25Leu a -
ian , and ha he wo gene ic al e a ions we e ansmi ed
in sepa a e alleles. Mo e impo an ly, we showed ha he
Se 80Ile mu a ion, bu no he P o25Leu a ian co-seg e-
ga ed wi h he VHL ype 2 pheno ype and ha he
P o25Leu a ian ne e occu ed in amily membe s who
had mani es a ions o he disease. Howe e , one amily
membe who had bo h gene ic al e a ions and one mem-
be wi h he Se 80Ile mu a ion wi hou he P o25Leu a -
ian showed no mani es a ions o he disease, sugges ing
ha his a ian is no pa hogenic o VHL.
In o de o p edic he pa hological ole o he P o25Leu
and Se 80Ile a ian s, i s we cons uc ed a mul iple
sequence alignmen using e olu iona y dis an species. I
has been shown p e iously ha he addi ion o mo e dis-
an ly ela ed sequences is essen ial o a mo e accu a e
p edic ion (20). Howe e , in ou case his app oach
e ealed ha bo h a ian s migh be neu al, which is in
con as wi h ea lie clinical indings sugges ing he dis-
ease causing ole o he Se 80Ile bu no o he P o25Leu.
The Se 80Ile has been p e iously epo ed only in one
VHL pa ien wi hou clea cell enal ca cinoma [23], bu
se e al o he missense mu a ions a codon 80 ha e been
desc ibed. The Se 80Gly mu a ion has been de ec ed in a
12-y -old pa ien wi h bila e al pheoch omocy omas and
mul iple congeni al mal o ma ions [24] and in a pa ien
wi h pheoch omocy oma [25]. The Se 80Asn mu a ion
was iden i ied in a Slo akian amily wi h VHL ype 1 phe-
no ype including panc ea ic isle umo in one a ec ed
indi idual [26]. In addi ion, he pa hogenic ole o
Se 80A g [27,28] and Se 80Asn mu a ions has been also
documen ed in VHL pa ien s [25]. Ou s udy shows o
he i s ime ha he Se 80Ile mu a ion is associa ed wi h
bila e al pheoch omocy oma p esen ed as a i s mani es-
a ion o he disease, which poin s ou ha VHL pa ien s
ha bo ing his mu a ion equi e ca e ul sc eening o phe-
och omocy oma.
The associa ion be ween a ious missense mu a ions a
codon 80 o he VHL gene and VHL disease in p e ious
s udies and ou s is no unexpec ed, since ou e olu iona y
alignmen analysis showed ha in his posi ion he Se is
a highly conse ed amino acid esidue among di e en
species. Mo eo e , compu a ional p o ein modeling o a
change o Se o Ile a posi ion 80 indica ed po en ially
impo an consequences on h ee-dimensional s uc u e
o he pVHL. Se 80 is a pola amino acid loca ed in he
hyd ophobic co e o he β-domain close o he α-β in e -
ace o pVHL and ElonginC in e ac ion. Acco ding o ou
p o ein modeling, he change o Se o a la ge and non-
Alignmen analysis o he VHL p o ein om Mus musculus (ENSMUSG00000033933), Ra us no egicus (ENSRNOG00000010258), Canis amilia is (ENSCAFG00000005149), Homo sapiens (ENST00000256474), Gallus gallus (ENSGALG000000013678), D osophila melanogas e (CG13221_CG13221-RA), Xenopus opicalis (ENSXET00000001448) and Taki ugu ub ipes (SINFRUG00000121189)Figu e 2
Alignmen analysis o he VHL p o ein om Mus musculus (ENSMUSG00000033933), Ra us no egicus
(ENSRNOG00000010258), Canis amilia is (ENSCAFG00000005149), Homo sapiens (ENST00000256474), Gallus gallus
(ENSGALG000000013678), D osophila melanogas e (CG13221_CG13221-RA), Xenopus opicalis (ENSXET00000001448)
and Taki ugu ub ipes (SINFRUG00000121189).

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pola Ile a amino acid posi ion 80 esul s in a complex
ea angemen o he h ee-dimensional s uc u e o he
p o ein binding in e ace, which dis u bs HIF1α and
ib onec in binding [29]. Mille and co-wo ke s ha e
al eady demons a ed ha VHL gene mu a ions wi hin L1
and L7 loops dis up he dynamic coupling o he pVHL
and HIF-1α and ha his may be an impo an mecha-
nism o he de elopmen o umo s [30]. O he s udies
using h ee-dimensional modeling and a compu e algo-
i hm FOLDEF p o ided a quan i a i e es ima ion o he
impo ance o he in e ac ions con ibu ing o he s abil-
i y o p o eins and p o ein complexes [31]. In he case o
he Se 80Ile, he ee ene gy di e ence be ween he wild
ype and mu an p o eins was 12.2 kcal/mol, which is
highe han he p e iously es ablished cu -o alue o he
de elopmen o clea -cell enal ca cinoma o pheoch o-
mocy oma [32].
Al hough mu a ions a ec ing he i s 53 codons o he
VHL gene ha e no e ec on he s uc u e o he sho e
VHL p o ein, a ew mu a ions a codons 25, 38, 46 and 52
ha e been conside ed o exe a pa hogenic ole. The
P o25Leu and Se 38P o ha e been iden i ied in pa ien s
wi h pheoch omocy omas, whe eas he Glu46S op and
Glu52Lys ha e been associa ed wi h VHL disease [32,27].
Howe e , Ro hbe g e al. epo ed a VHL pa ien wi h wo
VHL gene mu a ions, P o25Leu and P o86A g, in whom
he P o86A g was conside ed mo e likely o be pa hogenic
han he P o25Leu based on an allelic equency o 0.5%
o he la e a ian in anonymized DNA samples [14].
This P o25Leu a ian was de ec ed in o he heal hy VHL
Th ee-dimensional compu a ional modeling o he Se 80 and Ile80 esidues using CPK diag am (uppe pa ) and schema ic ep- esen a ion o hyd ogen-bonds be ween esidues Gly104 and Asn78, be ween A g79 and Ile151, be ween Se 80 and Ile151, beween Se 80 and P o103, be ween Se 80 and Leu153, and be ween P o103 and A g82 in wild ype pVHL and he conse-quences o he Se o Ile change a amino acid posi ion 80 (lowe pa )Figu e 3
Th ee-dimensional compu a ional modeling o he Se 80 and Ile80 esidues using CPK diag am (uppe pa )
and schema ic ep esen a ion o hyd ogen-bonds be ween esidues Gly104 and Asn78, be ween A g79 and
Ile151, be ween Se 80 and Ile151, beween Se 80 and P o103, be ween Se 80 and Leu153, and be ween P o103
and A g82 in wild ype pVHL and he consequences o he Se o Ile change a amino acid posi ion 80 (lowe
pa ). All igu es we e gene a ed wi h Swiss-PdbViewe in combina ion wi h POV-Ray.
BMC Medical Gene ics 2008, 9:29 h p://www.biomedcen al.com/1471-2350/9/29
Page 7 o 8
(page numbe no o ci a ion pu poses)
amily membe s om Poland and No h Ame ica [33,14].
In ag eemen wi h hese obse a ions, he P o25Leu ca -
ie in ou amily had no mani es a ions o he disease,
sugges ing ha his a ian does no ep esen a disease-
causing mu a ion.
Conclusion
The Se 80Ile mu a ion o he hl gene in a la ge Hunga ian
kind ed was ound o be associa ed wi h VHL ype 2 p e-
sen ing wi h bo h pheoch omocy oma and enal cell can-
ce . The e o e, pa ien s wi h his mu a ion equi e ca e ul
su eillance, including sea ch o pheoch omocy oma.
The P o25Leu a ian in his amily apea ed o be neu al,
bu i s co-exis ence made gene ic counseling di icul .
Abb e ia ions
VHL: on Hippel-lindau disease; pVHL: on Hippel-
Lindau p o ein; HIF: hypoxia inducible ac o .
Compe ing in e es s
The au ho (s) decla e ha hey ha e no compe ing in e -
es s.
Au ho s' con ibu ions
Acquisi ion o da a: AP, KB, PG, FF pe o med he molec-
ula biological analysis, AP, FF and IL pa icipa ed in
alignmen analysis and in h ee dimensional p o ein
modeling, MT and KR ob ained he clinical da a. Analysis
and in e p e a ion o da a: AP, FF, IL, MT and KR. D a ing
o he manusc ip : AP, KB, PG, MT, IL and KR. C i ical
e ision o he manusc ip o impo an in ellec ual con-
en : AP, IL, MT and KR. Adminis a i e, echnical, o
ma e ial suppo : AP, PG, KB, FF, IL, MT and KR. S udy
supe ision: KR.
Acknowledgemen s
W i en consen o publica ion was ob ained om he pa ien o hei el-
a i e. Suppo ed by g an s om he Hunga ian Minis y o Public Heal h
(EET 090/2006)
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