LCN6, a novel human epididymal lipocalin.
BACKGROUND
The lipocalin (LCN) family of structurally conserved hydrophobic ligand binding proteins is represented in all major taxonomic groups from prokaryotes to primates. The importance of lipocalins in reproduction and the similarity to known epididymal lipocalins prompted us to characterize the novel human epididymal LCN6.


RESULTS
LCN6 cDNA was identified by database analysis in a comprehensive human library sequencing program. Macaca mulatta (rhesus monkey) cDNA was obtained from an epididymis cDNA library and is 93% homologous to the human. The gene is located on chromosome 9q34 adjacent LCN8 and LCN5. LCN6 amino acid sequence is most closely related to LCN5, but the LCN6 beta-barrel structure is best modeled on mouse major urinary protein 1, a pheromone binding protein. Northern blot analysis of RNAs isolated from 25 human tissues revealed predominant expression of a 1.0 kb mRNA in the epididymis. No other transcript was detected except for weak expression of a larger hybridizing mRNA in urinary bladder. Northern hybridization analysis of LCN6 mRNA expression in sham-operated, castrated and testosterone replaced rhesus monkeys suggests mRNA levels are little affected 6 days after castration. Immunohistochemical staining revealed that LCN6 protein is abundant in the caput epithelium and lumen. Immunofluorescent staining of human spermatozoa shows LCN6 located on the head and tail of spermatozoa with the highest concentration of LCN6 on the post-acrosomal region of the head, where it appeared aggregated into large patches.


CONCLUSIONS
LCN6 is a novel lipocalin closely related to Lcn5 and Lcn8 and these three genes are likely products of gene duplication events that predate rodent-primate divergence. Predominant expression in the epididymis and location on sperm surface are consistent with a role for LCN6 in male fertility.
Hamil
Katherine G
KG
Department of Pediatrics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599, USA. khamil@med.unc.edu
Liu
Qiang
Q
Sivashanmugam
P
P
Anbalagan
M
M
Yenugu
Suresh
S
Soundararajan
Rama
R
Grossman
Gail
G
Rao
A J
AJ
Birse
Charles E
CE
Ruben
Stephen M
SM
Richardson
Richard T
RT
Zhang
Yong-Lian
YL
O'Rand
Michael G
MG
Petrusz
Peter
P
French
Frank S
FS
Hall
Susan H
SH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002899:Chromosomes, Human, Pair 9 / Q000235:genetics; D003001:Cloning, Molecular; D004822:Epididymis / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D054834:Lipocalins; D008253:Macaca mulatta / Q000235:genetics; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism
D002352:Carrier Proteins; C497171:LCN6 protein, human; D054834:Lipocalins; D012333:RNA, Messenger; D011994:Recombinant Proteins