Coltagen Biosynthesis: A Mini-Review Cluster

KARl I. KIVIRIKKO
Collagen Research Unit, Biocenter and Department of Medical Biochemistry, University of Oulu, Oulu,
Finland

Introduction
Collagen biosynthesis involves an unusually large
number of cotranslational and post-translational events,
many of which are unique to collagens and a few other
proteins with collagen-like amino acid sequences. The
intracellular events are catalyzed by five specific enzymes, some of which have further isoenzymes. Prolyl
4-hydroxylase plays a central role among these enzymes,
as the 4-hydroxyproline residues formed in its reaction
are essential for the folding of the newly synthesized collagen polypeptide chains into triple-helical molecules.
This enzyme will be discussed in the first article of the
series.
The four other specific intracellular enzymes of collagen synthesis will not be reviewed here. Lysyl hydroxylase has now been cloned from human (Hautala et al.,
1992; Yeowell et al., 1992), rat (Armstrong and Last,
199.5) and chick (Myllyl/i et al., 1991) sources, and the
catalytic properties of a recombinant enzyme expressed
in insect cells have been shown to be virtually identical
to those of an enzyme isolated from vertebrate tissues
(Armstrong and Last, 199.5; Krol et al., 1996; Pirskanen
et al., 1996). An isoenzyme termed lysyl hydroxylase
2 has very recently been cloned from human sources
(Valtavaara et al., 1997). Many mutations in the gene
for lysyl hydroxylase 1 have been characterized in families with Ehlers-Danlos syndrome type VI (Hyland et al.,
1992; Hautala et al., 1993; Ha et al., 1994; Pousi et al.,
1994; Heikkinen et al., 1997). Prolyl 3-hydroxylase and
the two collagen glycosyltransferases have been characterized in the past, but these enzymes have not yet been
cloned, and essentially no new data are available on
their properties.
Important new information is available on molecular
recognition in procollagen chain assembly, as will be disMatrix Biology Vol. 16/1997/98, pp. 3 5 5 - 3 5 6
1998 by Gustav Fischer Verlag

cussed in the second review of this series. New information is also available on Hsp47, a chaperone that may be
specifically involved in the assembly and/or secretion of
collagens, as will be reviewed in the third article. Nevertheless, this chaperone does not seem to be essential and
may possibly be replaced by others, as the assembly of
triple-helical procollagen molecules in insect cells (Lainberg et al., 1996; Myllyharju et al., 1997; Tomita et al.,
1997) and yeasts (Vuorela et al., 1997) does not require
any recombinant Hsp47.
The extracellular events in the biosynthesis of a fibril
forming collagen include cleavage of the N and C
propeptides, self-assembly into fibrils, and cross-link
formation. The two last reviews in this series discuss the
N and C proteinases, and lysyl oxidase and crosslink
formation.

References
Armstrong, L.C. and I,ast, J.A.: Rat lysyl hydroxylase: molecular cloning, mRNA distributions and expression in a baculovirus system. Biochim. Biophys. Acta 1264: 93-102, 1995.
Ha, V.T., Marshall, M.K., Elsas, L.J., Pinnell, S.R. and Yeowell,
H.N.: A patient with Ehlers-Danlos syndrome type VI is a
compound heterozygote for mutations in the lysyl hydroxylase gene. J. Clin. Invest. 93:1716-1721, 1994.
Hautala, T., Byers, M.G., EddB R.L., Shows, T. B., Kivirikko,
K.I. and Myllyl/i, R.: Cloning of human lysyl hydroxylase.
Complete cDNA-derived amino acid sequence and assignment of the gene to chromosome lp 36.2-36.3. Genomics
13: 62-69, 1992.
Hautala, T., Heikkinen, J., Kivirikko, K.I. and Myllyl/i, R.: A
large duplication in the gene for lysyl hydroxylase accounts
for the type VI variant of the Ehlers-Danlos syndrome in two
siblings. Genomics 15: 399-404, 1993.
Heikkinen, J., Toppinen, T., Yeowell, H., Krieg, T., Steinmann,
B., Kivirikko, K.I. and Myllyl/i., R.: Duplication of seven
exons in the lysyl hydroxylase gene is associated with longer
forms of a repetitive sequence within the gene and is acom-

356

K . I . Kivirikko

mon cause for the type VI variant of Ehlers-Danlos syndrome. Am. J. Hum. Genet. 60: 48-56, 1997.
Hyland, J., Ala-Kokko, L., Royce, P., Steinmann, B., Kivirikko,
K.I. and Myllyl/i, R.: A homozygous stop codon in the lysyl
hydroxylase gene in two siblings with Ehlers-Danlos syndrome type VI. Nat. Genet. 2: 228-231, 1992.
KroI, B.J., Murad, S., Walker, L.C., Marshall, M.K., Clark,
W.L., Pinnell, S.R. and Yeowell, H.N.: The expression of a
functional, secreted human lysyl hydroxylase in a baculovirus system. J. Invest. Dermatol. 106:1 l-16, 1996.
Lamberg, A., Helaakoski, T., Myllyharju, J., Peltonen, S., Notbohm, H., Pihlajaniemi, T. and Kivirikko, K.I.: Characterization of human type III collagen expressed m a baculovirus
system. Production of a protein with a stable triple helix requires coexpression with the two types of recombinant prolyl
4-hydroxylase subunit. J. Biol. Chem. 271: 11988-11995,
1996.
Myllyharju, J., Lamberg, A., Notbohm, H., Fietzek, P.P., Pihlajaniemi, T. and Kivirikko, K.I.: Expression of wild-type and
modified proc~ chains of human type I procollagen in insect
cells leads to the formation of stable [c(l(l)L,c(2(I) heterotrimers and [o:1(I)]~ homotrimers but not 10~2(I)1~ homotrimers. J. Biol. Chem. 272:21824-21830, 1997.
MyllylS, R., Pihlajaniemi, T., Pajunen, L., Turpeenniemi-Hujanen, T. and Kivirikko, K.I.: Molecular cloning of chick lysyl
hydroxylase. Little homology in primary structure to the two
types of subunit of prolyl 4-hydroxylase..I. Biol. Chem. 266:
2805-2810, 1991.
Pirskanen, A., Kaimio, A.-M., Myllyl/i, R. and Kivirikku, K.I.:
Site-directed mutagenesis of human lysyl hydroxylase expressed in insect cells. Identification of histidine residues and

an aspartic acid residue critical for catalytic activity. J. Biol.

Chem. 271: 9398-9402, 1996.
Pousi, B., Hautala, T., Heikkinen, J., Pajunen, L., Kivirikko,
K.I. and Myllyl/i, R.: Alu-Alu recombination results in a duplication of seven exons in the lysyl hydroxylase gene in a patient with the type VI variant of Ehlers-Danlos syndrome.
Am..1. Hum. Genet..55: 899-906, 1994.
Tomita, M., Kitajima, T. and Yoshizato, K.: Formation of recombinant human procollagen I heterotrimers in a bac
ulovirus expression system. I. Biochem. 121: 1061-1069,
1997.
Valtavaara, M., Papponen, H., Pirttil/i, A.-M., Hiltunen, K.,
Helander, H. and Myllyl/i, R.: Cloning and characterization
of a novel lysyl hydroxylase isoform highly expressed in pancreas and muscle. J. Biol. Chem. 272: 6831-6834, 1997.
Vuorela, A., Myllyharju, J., Nissi, R., Pihlajaniemi, T. and
Kivirikko, K.I.: Assembly of human prolyl 4-hydroxylase and
type III collagen in the yeast Pichia pastoris: formation of a
stable enzyme tetramer requires coexpression with collagen
and assembly of a stable collagen requires coexpression with
prolyl 4-hydroxylase. EMBO J. 16: 6702-67t 2, 1997.
Yeowell, H.N., Ha, V., Walker, L C , Murad, S. and Pinnell,
S.R.: Characterization of a partial cDNA for lysyl hydroxylase from human skin fibroblasts; lysyl hydroxylase mRNAs
are regulated differently by minoxidil derivatives and hydralazine. J. Invest. Dermatol. 99: 864-869, 1992.
Dr. Kari I. Kivirikko, Department of Medical Biochemistry,
University of Oulu, FIN-90220 Oulu, Finland
Received October 28, 1997