Molecular cloning of PARP (proline/arginine‐rich protein) from human cartilage and subsequent demonstration that PARP is a fragment of the NH₂‐terminal domain of the collagen α2(XI) chain

Abstract

We report the molecular cloning of a proline/arginine-rich protein (called PARP) from human cartilage using the polymerase chain reaction (PCR) and degenerate oligonucleotides based on the previously published amino acid sequence of bovine PARP [l]. Subsequently, a reverse transcriptionpolymerase chain reaction (RT-PCR) was performed with poly(A)-rich RNA from human cartilage using a sense oligonucleotide derived from PARP and an anti-sense oligonucleotide derived from the known sequence of the human collagen a2(XI) chain [2]. Nucleotide sequencing of the PCR product demonstrated that PARP is a fragment of the NHz-terminal non-collagenous (NC3) domain of the collagen a2(XI) chain.

Diversity in the processing events at the N‐terminus of type‐V collagen

“…This peptide sequence is located at the beginning of the variable region which, with the constant region defined by Zhidkova et al (1993), is in the acidic subdomain of the pro-al(V) chain (Greenspan et al, 1991). The variable region is situated after the prolinehrginine-rich-protein-like domain, which is the most N-terminal region of the molecule (see Fig.…”

“…As shown in Fig. 2, the specificity of antibody binding is ascertained by the fact that these antibodies failed to recognize an unrelated peptide (multiple-antigen peptide XI) and that the preimmune sera did not exhibit any reactivity to the Zhidkova et al (1993). The human cDNA-derived amino acid sequence of pro-cxl(V) is shown from the beginning of the acidic subdomain (Greenspan et al, 1991) peptides.…”

Characterization of Two Genes Coding for a Similar Four‐Cysteine Motif of the Amino‐Terminal Propeptide of a Sea Urchin Fibrillar Collagen

“…Drosophiln sog and Xenopus chordin proteins contain four repeats of a cysteine-rich motif which is distantly related to the cysteine-rich globular region of the fibrillar procollagen chains and the thrombospondin molecules (Francois et al, 1995;Sasai et al, 1995). The basic part of the long globular region present in the N-propeptide of structure 111 is also called the thrornbospondin 1 (tsp-I) motif (Bork, 1992) or the PARP domain (prolinehrginine-rich protein ; Zhidkova et al, 1993). The PARP domain of the fibrillar pro-cxl (V), pro-crl(X1) and pro-n2(XI) chains (Zhidkova et al, 1993;Greenspan et al, 1991;Takahara et al, 1991: Yoshioka andRamirez, 1990) is also present in the non-fibrillar collagen types IX, XII, XIV and XVIIl (Rehn and Pihlajaniemi, 1994) and in thrombospondin molecules (Lawler and Hynes, 1986;Adams and Lawler, 1993).…”

“…The basic part of the long globular region present in the N-propeptide of structure 111 is also called the thrornbospondin 1 (tsp-I) motif (Bork, 1992) or the PARP domain (prolinehrginine-rich protein ; Zhidkova et al, 1993). The PARP domain of the fibrillar pro-cxl (V), pro-crl(X1) and pro-n2(XI) chains (Zhidkova et al, 1993;Greenspan et al, 1991;Takahara et al, 1991: Yoshioka andRamirez, 1990) is also present in the non-fibrillar collagen types IX, XII, XIV and XVIIl (Rehn and Pihlajaniemi, 1994) and in thrombospondin molecules (Lawler and Hynes, 1986;Adams and Lawler, 1993). More recently, different isoforms of the chicken and rat pro-(11 (XI) collagen chains (Zhidkova et al, 1995;Thom Oxford et al, 1995) and human and mouse proa2(XI) collagen chains (Zhidkova et al, 1995;Tsumaki and Kimura.…”

“…The pro-a2(1) chains and the alternatively spliced form of the pro-nl(1I) chains represented the N-propeptide configuration of structure I T (Vuorio and de Cromhrugghe, 1990;Ryan and Sandell, 1990). The N-propeptide of structure 111 included the pro-al(V), pro-al(X1) and pro-rx2(X1) chains (Greenspan et al, 1991;Takahara et al, 1991 ;Yoshioka and Ramirez, 1990;Zhidkova et al, 1993). a complex alternative splicing system which involves the sequence coding for the acidic subdomain of the N-propeptide.…”